Broad RNA Interference−Mediated Antiviral
Immunity and Virus-Specific Inducible
Responses in Drosophila
This information is current as
of June 18, 2017.
Cordula Kemp, Stefanie Mueller, Akira Goto, Vincent
Barbier, Simona Paro, François Bonnay, Catherine Dostert,
Laurent Troxler, Charles Hetru, Carine Meignin, Sébastien
Pfeffer, Jules A. Hoffmann and Jean-Luc Imler
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J Immunol 2013; 190:650-658; Prepublished online 19
December 2012;
doi: 10.4049/jimmunol.1102486
http://www.jimmunol.org/content/190/2/650
The Journal of Immunology
Broad RNA Interference–Mediated Antiviral Immunity and
Virus-Specific Inducible Responses in Drosophila
Cordula Kemp,*,1 Stefanie Mueller,*,1,2 Akira Goto,* Vincent Barbier,* Simona Paro,*
François Bonnay,* Catherine Dostert,* Laurent Troxler,* Charles Hetru,* Carine Meignin,*
Sébastien Pfeffer,† Jules A. Hoffmann,* and Jean-Luc Imler*,‡
V
iruses represent an important class of pathogens, causing
serious concern for human health, as well as important
economic losses in crops and animals. Because they
replicate inside cells, and rely for the most part on host cell molecular machineries for their replication, viruses pose specific
challenges to the immune system. Two major strategies of antiviral
*CNRS-UPR9022, Institut de Biologie Moléculaire et Cellulaire, 67084 Strasbourg
Cedex, France; †CNRS-UPR9002, Institut de Biologie Moléculaire et Cellulaire;
67084 Strasbourg Cedex, France; and ‡Faculté des Sciences de la Vie, Université
de Strasbourg; 67083 Strasbourg Cedex, France
1
C.K. and S.M. contributed equally to this work.
2
Current address: Bernhard-Nocht-Institut for Tropical Medicine, Molecular Entomology, Hamburg, Germany
Received for publication August 29, 2011. Accepted for publication November 5,
2012.
This work was supported by the National Institutes of Health (PO1 AI070167), the
Agence Nationale de la Recherche (ANR-09-MIEN-006-01), the Balzan Foundation
(to J.A.H.), the European Research Council (ERC Starting Grant ncRNAVIR 260767
to S.P.), the Investissement d’Avenir Program Laboratoire d’Excellence (NetRNA
ANR-10-LABX-36), and the Centre National de la Recherche Scientifique.
The sequences presented in this article have been submitted to the Gene Expression
Omnibus (http://www.ncbi.nlm.nih.gov/geo/) under accession number GSE31542
and to the National Center for Biotechnology Information Small Read Archive
(http://www.ncbi.nlm.nih.gov/sra) under accession number GSE41007.
Address correspondence and reprint requests to Prof. Jean-Luc Imler, IBMC-CNRS/
Université de Strasbourg, Rue Rene Descartes, Strasbourg, Alsace 67084, France.
E-mail address: [email protected]
The online version of this article contains supplemental material.
Abbreviations used in this article: AGO, Argonaute; CrPV, cricket paralysis virus;
Dcr-2, Dicer-2; DCV, Drosophila C virus; dpi, day postinfection; DXV, Drosophila X
virus; FHV, Flock House virus; IIV-6, invertebrate iridescent virus type 6; Imd,
immune deficiency; MEKK1, MEK kinase 1; RNAi, RNA interference; SINV, Sindbis virus; siRNA, small interfering RNA; TotM, Turandot M; Upd, unpaired; VSV,
vesicular stomatitis virus.
Copyright Ó 2013 by The American Association of Immunologists, Inc. 0022-1767/13/$16.00
www.jimmunol.org/cgi/doi/10.4049/jimmunol.1102486
resistance have been described. In mammals, viral infection is first
detected by pattern recognition receptors of the Toll- and RIG-I–
like families that sense the viral nucleic acid and trigger the induction of IFNs and other cytokines (1). These factors activate
the production of antiviral molecules, such as protein kinase R
or oligo-29, 59-adenylate synthetase, that contain the infection and
contribute to the activation of the adaptive immune response (2).
In plants, viral nucleic acids are recognized by enzymes of the
Dicer family, which produce small interfering RNAs (siRNAs) of
21–24 nucleotides. These siRNAs are then loaded onto molecules
of the Argonaute (AGO) family and will guide them toward RNAs
with complementary sequences; targeted RNAs are then either
sliced by AGO, or their translation is inhibited. This RNA interference (RNAi) mechanism provides efficient and sequencespecific protection against viral infections (3).
RNAi also plays an important role in the control of viral infections in insects, as shown by the production of virus-derived
siRNAs in infected flies, and the increased susceptibility to viral
infection of Drosophila mutants for the genes Dcr-2 and AGO2
(3–6). In addition, several reports indicate that an inducible response also contributes to the control of viral infections (7–15).
We previously showed that infection with Drosophila C virus
(DCV), a member of the Dicistroviridae family, leads to induction
of some 130 genes (11). Analysis of the regulation of one of these
genes, vir-1, revealed the presence of functionally important
binding sites for the transcription factor STAT in its promoter. The
induction of vir-1, as well as several other DCV-induced genes,
was found to be dependent on the gene hopscotch (hop), which
encodes the only JAK kinase in Drosophila. Furthermore, hop
mutant flies succumb more rapidly than do wild-type controls,
with a higher viral load, to DCV infection (11). The Toll and
immune deficiency (Imd) pathways, initially characterized for
their role in the control of bacterial and fungal infections, were
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The fruit fly Drosophila melanogaster is a good model to unravel the molecular mechanisms of innate immunity and has led to
some important discoveries about the sensing and signaling of microbial infections. The response of Drosophila to virus infections
remains poorly characterized and appears to involve two facets. On the one hand, RNA interference involves the recognition and
processing of dsRNA into small interfering RNAs by the host RNase Dicer-2 (Dcr-2), whereas, on the other hand, an inducible
response controlled by the evolutionarily conserved JAK-STAT pathway contributes to the antiviral host defense. To clarify the
contribution of the small interfering RNA and JAK-STAT pathways to the control of viral infections, we have compared the
resistance of flies wild-type and mutant for Dcr-2 or the JAK kinase Hopscotch to infections by seven RNA or DNA viruses
belonging to different families. Our results reveal a unique susceptibility of hop mutant flies to infection by Drosophila C virus and
cricket paralysis virus, two members of the Dicistroviridae family, which contrasts with the susceptibility of Dcr-2 mutant flies to
many viruses, including the DNA virus invertebrate iridescent virus 6. Genome-wide microarray analysis confirmed that different
sets of genes were induced following infection by Drosophila C virus or by two unrelated RNA viruses, Flock House virus and
Sindbis virus. Overall, our data reveal that RNA interference is an efficient antiviral mechanism, operating against a large
range of viruses, including a DNA virus. By contrast, the antiviral contribution of the JAK-STAT pathway appears to be virus
specific. The Journal of Immunology, 2013, 190: 650–658.
The Journal of Immunology
Materials and Methods
Fly strain culture and infection
Oregon-R (OR) and yw were used as wild-type control flies. The hopM38/msv1,
Dcr-2L811fsX, and Dcr-2R416X mutant flies were previously described (17–
19). A genomic rescue of the Dcr-2 gene was established with the Fosmid
FlyFos017074 (transgeneome.mpi-cbg.de) inserted at the landing site
attP40 (2L), and the transgenic chromosome was recombined with the
deficiency Df(2R)BSC45, which uncovers the Dcr-2 locus. For the rescue
experiments, Dcr-2 mutants were crossed with the deficiency Df(2R)
BSC45 or the Df(2R)BSC45–Dcr-2 rescue line. Flies were fed on standard
cornmeal–agar medium at 25˚C. All fly lines were tested for Wolbachia
infection and cured whenever necessary. Viral stocks were prepared in 10
mM Tris-HCl, pH 7.5, with the exception of vesicular stomatitis virus
(VSV), which was used directly from Vero cell culture supernatant [VSV
4 3 109 PFU/ml; DCV 5 3 1010 PFU/ml; CrPV 1 3 109 PFU/ml; FHV
5.5 3 109 PFU/ml; DXV 4.4 3 107 PFU/ml, invertebrate iridescent virus
type 6 (IIV-6) 4.4 3 1011 PFU/ml; and SINV 5 3 108 PFU/ml]. Infections
were performed with 4- to 6-d-old adult flies by intrathoracic injection
(Nanoject II apparatus; Drummond Scientific) with viral particles, indicated in the figure legends. Injection of the same volume (4.6 nL) of 10
mM Tris-HCl, pH 7.5, was used as a control. For bacterial infection, flies
were pricked with a thin needle previously dipped in a concentrated
overnight culture of Escherichia coli and Micrococcus luteus in Luria–
Bertani medium. Infected flies were then incubated at room temperature,
or at 29˚C in the case of hopM38/msv1 and the corresponding control flies,
and monitored daily for survival, or frozen for RNA isolation and virus
titration at the indicated time points.
Cell culture and virus titration
Vero R cells were grown in DMEM (Invitrogen) supplemented with 10%
FCS (Biowest), penicillin/streptomycin (Invitrogen), nonessential amino
acid mix (Invitrogen), 10 mM pyruvate (Life Technologies), and 200 mM Lglutamine (Invitrogen). Kc167 and S2 cells were grown in Schneider’s
medium (Biowest) supplemented with 10% FCS, GlutaMAX (Invitrogen),
and penicillin/streptomycin (1003 mix, 10 mg/ml/10,000 U; Invitrogen).
VSV and SINV were titrated from infected flies by plaque assay on Vero R
cells. DCV, CrPV, FHV, and IIV-6 were titrated on Kc167 (DCV, CrPV,
and FHV) or S2 (IIV-6) cells by the Reed–Muench method to calculate
50% tissue culture–infective dose and converted to PFU with a conversion
factor of 0.7.
RNA analysis
Total RNA from infected flies was isolated using TRI Reagent RT bromoanisole solution (MRC), according to the manufacturer’s instructions.
Total RNA, 1 mg, was reverse transcribed using iScript cDNA Synthesis
Kit (Bio-Rad). The reverse transcription was run in the T3000 Thermocycler (Biometra), with the following PCR program: step 1: 65˚C for 5
min, step 2: 4˚C for 5 min, step 3: 25˚C for 10 min, step 4: 42˚C for 60
min, and step 5: 70˚C for 15 min. A total of 100 ng cDNA was used for
quantitative real-time PCR, using the iQ Custom SYBR Green Supermix
Kit (Bio-Rad). The PCR was performed using the CFX384 Real-Time
System (Bio-Rad) with the following program: step 1: 95˚C for 3 min,
step 2: 95˚C for 10 s, step 3: 55˚C for 30 s, repeated 39 times from step 2.
Primers used for qPCR were as follows: RpL32 (forward 59-GACGCTTCAAGGGACAGTATCTG-39; reverse 59-AAACGCGGTTCTGCATGAG-39),
vir-1 (forward 59-GATCCCAATTTTCCCATCAA-39; reverse 59-GATTACAGCTGGGTGCACAA-39), drosomycin (forward 59-CGTGAGAACCTTTTCCAATATGATG-39; reverse 59-TCCCAGGACCACCAGCAT-39), and
diptericin (forward 59-GCTGCGCAATCGCTTCTACT-39; reverse 59TGGTGGAGTGGGCTTCATG-39). Turandot M (TotM), upd, upd2, and
upd3 expression levels were quantified using the Brilliant II QRT-PCR
Core Reagent Kit, 1-step (Stratagene). The reaction took place in a total
volume of 20 ml, using the Taqman Gene Expression Assay [TotM
(Dm02362087 s1), upd (os) (Dm01843792_g1), upd2 (Dm01844134 g1),
upd3 (custom-designed upd3exon2-ANY), and RpL32 (Dm02151827 g1),
all from Applied Biosystems]. We used the 7500 Fast Real-Time PCR
System (Applied Biosystems) with following PCR program: step 1: 45˚C for
30 min, step 2: 95˚C for 10 min, step 3: 95˚C for 15 s, step 4: 60˚C for
1 min, repeated 39 times from step 3. In all cases, gene expression was
normalized to the ribosomal protein gene RpL32.
For IIV-6, the expression of the annotated genes 206R, 224L, 244L, and
261R was assessed by strand-specific RT-PCR. We used SuperScript III
Reverse Transcriptase specifically adapted for gene-specific priming and
followed the manufacturer’s protocol (Invitrogen). Briefly, primer pairs
were designed to amplify regions of the IIV-6 genome exhibiting or not
exhibiting a high density of small RNA reads. Total RNA,1 mg, extracted
from infected S2 cells was reverse transcribed with 2 pmol of either forward (F) or reverse (R) primer and 200 U of SuperScript III Reverse
Transcriptase. The reaction was then incubated for 1 h at 55˚C. Then 1 ml
of the resulting cDNA was used to perform 25 cycles of PCR, using Taq
DNA polymerase (Invitrogen) and both F and R primers. The primer pairs
were as follows: 206R (forward: 59-AAGGAAAGTGGCGAGTACGA-39,
reverse 59-AACAAACCCGTTTTCTTCCA-39); 224L (forward: 59-CCACCATCACATTGACCTTG-39, reverse: 59-ATAAGCGAACCCGAAATCA-39);
244L (forward: 59-TGGAAAAGAGTGGTCCCATTT-39, reverse: 59-TGTACCTCCCGGAAGATTT-39); 261R (forward: 59-CAGCCCCATCCGAATTACTA-39, reverse: 59-CTGCAACTGCAGAAATTTGA-39). The PCR bands
were sequenced to verify their viral origin.
Statistical analysis
An unpaired two-tailed Student t test was used for statistical analysis of
data with GraphPad Prism (GraphPad Software). The p values , 0.05 were
considered statistically significant. Survival curves were plotted and analyzed by log-rank analysis (Kaplan–Meier method) using GraphPad Prism
(GraphPad Software).
DNA microarray analysis
For each sample, Tris-injected, DCV-infected (11), and FHV- and SINVinfected, three biologically independent samples comprising 45 male
Oregon-R flies were used. RNA extraction, biotinylation, and hybridization
to Affymetrix Drosophila GeneChip microarrays (Affymetrix) were performed as described (20). The Affymetrix Microarray Suite 5.0 (Affymetrix) or Excel (Microsoft) with a combination of built-in functions and
custom formulae was used for data analysis. Raw data were sorted with the
“absent-marginal-present flags” generated by the Microarray Suite functions. Although an absent flag might indicate that no mRNA of a particular
type was present in a sample, marginal flags and absent flags may indicate
problems with the hybridization; therefore, only data points marked as
present in at least one replicate were retained. The remaining data mass
for each microarray was then normalized to itself, making 1 the median
of all the measurements. A gene was considered induced if present in at
least one replicate, with a virus/Tris ratio higher than 2 for at least one of
the time points. Classification of gene functions was analyzed by David
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also thought to play a part in the control of viral infections.
Whereas the Toll pathway was associated with resistance to the
Drosophila X virus (DXV) (15), the Imd pathway was implicated
in the control of Sindbis virus (SINV) (7) and cricket paralysis
virus (CrPV) (9).
Altogether, the data in the present literature point to the involvement of both RNAi and an inducible expression of effector
molecules to counter viral infections in insects (5, 16). However,
whereas RNAi was shown to contribute to resistance to several
RNA viruses (with either single-stranded genomes of both polarities or double-stranded genomes), most studies on the inducible
response have so far focused on a single virus. As a result, the
global significance of the inducible response for the control of
viral infections remains poorly understood. In particular, it is
unclear at present if the JAK-STAT pathway is involved in a
general antiviral response, providing broad antiviral immunity, or
if it acts specifically on a critical step in the replication cycle of a
specific virus or virus family. To address this important question,
we have compared the resistance of a mutant for the JAK-STAT
pathway to infection by seven RNA or DNA viruses. We find that
hop mutant flies are more susceptible than wild-type controls to
infections by the Dicistroviridae DCV and CrPV, but exhibit either
no or a weak phenotype for other viruses, suggesting that the
JAK-STAT pathway–dependent inducible response is virus specific. Genome-wide transcript profiling shows that infection by
two other RNA viruses, Flock House virus (FHV; Nodaviridae)
and SINV (Alphaviridae), leads to upregulation of $400 genes,
which only partially overlap with those induced by DCV. Overall,
our data indicate that the siRNA pathway exerts broad antiviral
activity and affects both RNA and DNA viruses, with virusspecific inducible responses contributing to the control of viral
infections in Drosophila.
651
652
Bioinformatics Resources 6.7 (21). The data set for FHV and SINV was
submitted to the Gene Expression Omnibus database (http://www.ncbi.
nlm.nih.gov/geo/) with the accession number GSE31542.
Assembly, sequencing, and analysis of small RNA libraries
The small RNA library of S2 cells and whole flies was constructed as
described (22) and sequenced by the Illumina 2G Analyzer. Reads were
then aligned to a reference consisting of the IIV-6 genome from the National Center for Biotechnology Information (accession code NC_003038)
using the Bowtie program with standard parameters in genome assembly.
Reads aligning to the IIV-6 genome with a maximum of one mismatch were
retained and analyzed using in-house Perl scripts and Excel. Sequences were
submitted to the National Center for Biotechnology Information Small Read
Archive (http://www.ncbi.nlm.nih.gov/Traces/sra/sra.cgi?) under the accession number GSE41007.
Results
RNAi provides broad antiviral protection in Drosophila
which is characteristic of the accumulation of iridescent viral
particles, than in wild-type controls (Fig. 1A). Dcr-22/2 flies were
significantly more susceptible to IIV-6 infection than were the
corresponding wild-type (Fig. 1B). A fraction of Dcr-22/2 flies
injected with buffer also died in the course of the experiment,
confirming the increased sensitivity to stress associated with
mutations of the siRNA pathway (31). The decreased survival
time correlated with a 20-fold increased viral load in Dcr-2 mutant
flies at 10 d postinfection (dpi) (Fig. 1C). Similar results were
obtained when a different null allele of Dcr-2 was used, and the
IIV-6 susceptibility phenotype was rescued by a wild-type genomic Dcr-2 transgene (Fig. 1D). The r2d22/2 and AGO22/2 null
mutant flies also exhibited increased sensitivity to IIV-6 (Fig.
1E). AGO22/2 flies contained more viral DNA than did wild-type
controls, confirming that this gene participates in the control of
infection (Fig. 1F).
We next sequenced small RNA libraries prepared from IIV-6–
infected S2 cells or adult flies. We observed several hundreds of
thousands of reads matching the IIV-6 genome in both infected S2
cells and wild-type flies, but not in control noninfected S2 cells
(Supplemental Table I). The large majority of these reads had a
size of 21 nucleotides, which is characteristic for processing by
the RNase Dicer-2 (Dcr-2). This peak was absent from the library
prepared from infected Dcr-22/2 mutant flies (Fig. 2A). These
data indicate that Dcr-2 generates 21-nucleotide IIV-6–derived
siRNAs in infected flies, and raise the question of the nature of the
substrate used by Dcr-2 in the context of this infection. As previously reported for RNA viruses, the number of reads matching
FIGURE 1. Dcr-2 is involved in host defense against the DNA virus IIV-6. (A) Upon injection of IIV-6 (5000 PFU) in wild-type (yw) and Dcr-2R416X
mutant flies, typical blue paracrystalline structures appeared earlier in the abdomen (arrowhead) of the mutant flies. Representative individuals 10 dpi are
shown. (B) Groups of 20 wild-type (yw) or Dcr-2R416X mutant flies were injected with IIV-6 or Tris, and survival was monitored daily. The difference
between the wild-type and Dcr-2 mutant flies is statistically significant. (C) Viral titer in groups of five wild-type (yw) or Dcr-2R416X mutant flies was
monitored 10 dpi. (D) Rescue of the hemizygous Dcr-2L811fsX for the IIV-6 susceptibility phenotype by a transposon expressing a wild-type Dcr-2
transgene. Dcr-2L811fsX hemizygous flies (Dcr-2L811fsX/Df) are significantly more susceptible than Dcr-2L811fsX hemizygous flies complemented by a wildtype Dcr-2 transgene (Dcr-2L811fsX/Df rescue). Df is Df(2R)BSC45, a deficiency that fully uncovers the Dcr-2 locus. All control and genomic rescued flies
are in CantonS background. (E) Survival rate of wild-type (yw), R2D21, and AGO2414 mutant flies upon IIV-6 or Tris injection. (F) IIV-6 DNA load was
determined by quantitative PCR in four groups of six flies of the indicated genotype at 10 dpi. For all panels, the data represent the mean and SD of at least
three independent experiments, and the difference between controls and mutant flies is statistically significant. *p , 0.05, ***p , 0.001. All experiments
are performed at 22˚C (A, C, F) or 25˚C (B, D, E).
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Several independent studies, including our own, have established
that RNAi, and more precisely the siRNA pathway, serves as an
efficient host defense against RNA viruses. These include viruses
with a single-stranded genome of both (+) and (2) polarity and
dsRNA viruses (23–30), and we confirmed that flies mutant for
Dcr-2 died more rapidly than wild-type controls when they were
infected with DCV, CrPV, FHV, SINV, VSV (Rhabdoviridae), and
DXV (Birnaviridae) (data not shown). Next, we addressed the
question whether the siRNA pathway also participated in the
control of a DNA virus infection, and infected wild-type and RNAi
mutant flies with IIV-6 (Iridoviridae). Infection of Dcr-2 mutant
flies led to a more rapid and intense appearance of blue color,
VIRUS-SPECIFIC INDUCIBLE RESPONSE IN DROSOPHILA
The Journal of Immunology
653
each strand of the viral genome was very similar (Supplemental
Table I). However, unlike RNA viruses, the virus-derived siRNAs
were not uniformly distributed along the viral genome. Rather,
several hotspots were observed, revealing that specific regions of
the viral genome generate the siRNAs (Fig. 2B, 2C). These peaks
do not correlate with the intensity of transcription of the viral
genome, and some highly transcribed regions are located in areas
not generating significant levels of siRNAs (32). The strong
symmetry of the peaks observed in S2 cells and wild-type flies
suggests that these regions are transcribed on both strands and
generate dsRNA. Indeed, we could detect bidirectional transcription in the areas of the viral genome covered by the peaks (Fig.
2D). By contrast, transcription of only one strand of the DNA
genome was detected for the locus 261R, which is located in
a region that does not produce significant amounts of siRNAs.
Overall, these results indicate that the siRNA pathway in Drosophila can also protect against a DNA virus infection.
The JAK kinase Hopscotch does not confer broad antiviral
immunity
To test the contribution of the JAK-STAT pathway in antiviral
immunity in Drosophila, we injected loss-of-function mutants of
the JAK kinase Hopscotch (hopM38/msv1) with different ssRNA,
dsRNA, and DNA viruses. As previously described, hopM38/msv1
mutant flies die more rapidly than do wild-type controls following
DCV infection, and contain ∼10-fold more virus (Fig. 3A). By
contrast, we did not observe significant differences in survival
between wild-type and hopM38/msv1 mutant flies upon infection
with the alphavirus SINV (Fig. 4A), and the viral titers 2 dpi were
not significantly different in wild-type and hopM38/msv1 mutant
flies (data not shown), indicating that the JAK-STAT pathway does
not contribute to resistance to this virus. The hopM38/msv1 mutant
flies, as well as wild-type flies, also resisted infections by the
rhabdovirus VSV and by the nodavirus FHV (Fig. 4B, 4C). A
slight reduction in survival was observed in the case of the dsRNA
virus DXV (Birnaviridae) and the DNA virus IIV-6 (Fig. 4D, 4E).
However, the difference between wild-type and hopM38/msv1 mutant
flies was only statistically significant in the case of DXV infection.
Furthermore, we did not observe statistically significant differences
in the DXV and IIV-6 viral titers in wild-type and hop M38/msv1
mutant flies in the format of our assays (data not shown).
Overall, our data indicate that the JAK-STAT pathway is critical for
host defense against DCV, but plays a minor role for DXV and IIV-6
and is essentially dispensable in the case of FHV, SINV, and VSV. We
therefore tested CrPV, another member of the Dicistroviridae family
known to infect Drosophila. We observed a decrease in survival
and a significant increase in viral titers in CrPV-infected hopM38/msv1
mutant flies compared with wild-type flies (Fig. 3B). In conclusion,
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FIGURE 2. Virus-derived siRNAs in S2 cells and Drosophila adult flies infected by the DNA virus IIV-6. RNA was extracted 5 dpi from S2 cells infected
by IIV-6 (MOI 0.01) and adult wild-type (yw) or mutant (Dcr-2R416X) flies injected with IIV-6 (5000 PFU per fly). (A) Size distribution of the small RNAs
matching the viral genome in S2 cells and adult flies of the indicated genotype. (B and C) Distribution of the 21-nucleotide siRNAs from the S2 cell (B) and
yw adult fly (C) libraries along the IIV-6 genome. Each IIV-6–derived small RNA is represented by the position of its first nucleotide. The IIV-6–derived
small RNAs matching the upper and lower strand of the DNA genome are respectively shown above (positive reads number) and below (negative reads
number) the horizontal axis, which represents the 212482bp genome. In (B), the number of reads for four peaks going off-scale is indicated next to them, in
italics. (D) Strand-specific RT-PCR with primers corresponding to the annotated viral genes 206R, 224L, 244L, and 261R. The experiment was performed in
the presence (+) or absence (2) of RT. NI, Noninfected; F and R, forward and reverse strand primer used for reverse transcription.
654
VIRUS-SPECIFIC INDUCIBLE RESPONSE IN DROSOPHILA
our data indicate that the JAK-STAT pathway in Drosophila confers
protection against some viruses—in particular, the Dicistroviridae—
but does not provide broad antiviral immunity.
Inducible gene expression in FHV- and SINV-infected flies
The above results raised the question of whether an inducible
response contributes to host defense against viruses other than
DCV and CrPV. We therefore conducted a genome-wide microarray analysis using Affymetrix DNA microarrays to monitor gene
expression in flies infected by FHV (2 and 3 dpi) or SINV (4 and 8
dpi), and compared the data with those obtained for DCV infection
(1 and 2 dpi). The time points for this analysis were chosen to take
into account the different kinetics of replication and colonization
of Drosophila by the different viruses (11, 24). For each virus, we
observed a large overlap between the genes induced at the first and
second time points. We then pursued our analysis, focusing on the
genes induced either at the first or at the second time point. The
microarray data revealed that 487 and 201 genes were induced or
FIGURE 4. Susceptibility of flies mutant for the JAK kinase Hopscotch to infection by SINV, VSV, FHV, DXV, and IIV-6. Groups of 20 wild-type
(OR) or hop mutant flies were injected with SINV (A), VSV (B), FHV (C), DXV (D), or IIV-6 (E), and survival was monitored. For VSV and SINV, the Tris
buffer control injection is also shown, because hop mutant flies exhibited decreased survival at 29˚C after day 16 upon both buffer and virus injection.
Kaplan–Meier analysis of the results of at least two independent experiments reveal a statistically significant difference in survival between wild-type and
hop mutant flies only in the case of DXV. *p , 0.05.
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FIGURE 3. The JAK kinase Hopscotch is involved in host defense against DCV and CrPV. (A and B) Groups of 20 wild-type (OR) or hopscotch (hopM38/msv1)
mutant flies were injected with DCV (500 PFU) (A) or CrPV (5 PFU) (B), and survival was monitored daily. The experiment was repeated three times, and
data represent the mean and SD. In the right panels, viral titer was determined in groups of five flies 2 dpi for DCV (A) and 1 dpi for CrPV (B). The data
represent the mean and SD of three independent experiments, and the difference between wild-type and hop mutant flies is statistically significant. *p ,
0.05, **p , 0.01, ***p , 0.001. (C) DCV and CrPV infection triggers induction of the genes upd2 and upd3, which encode cytokines activating the
JAK/STAT pathway. Flies were infected with DCV or CrPV, and expression of upd, upd2, and upd3 was monitored in groups of 10 flies at the indicated
time points by Taqman quantitative PCR. The results of at least two independent experiments are shown.
The Journal of Immunology
655
upregulated by a factor of at least 2 upon infection by FHV and
SINV, respectively. When analyzed with the same criteria, 166
genes were induced by DCV (Fig. 5A, Supplemental Table II).
The data of this transcriptomic analysis call for two comments.
First, we note that 42 genes were induced by all three viruses
(Fig. 5A). We compared this set of genes with microarray studies
performed on flies infected by fungi and bacteria (both extra- and
intracellular) to identify a potential signature specific for viral
infections (Supplemental Table III). We observed that a number of
genes, such as Frost, are upregulated similarly by all types of
infections, suggesting that they are induced by the stress of the
infection, rather than by recognition of specific characteristics
of the infecting microorganism. Of interest, other genes, such as
Vago, Obp99b, Mal-B1, Nmda1, CG8147, CG1572, l(2)gd1,
CG14906, CG10911, and Tsp42EI, appear to be induced only in
response to viral infections, and may represent the core of an
inducible antiviral gene expression program. The case of Obp99b
is particularly striking, as this gene is strongly upregulated by
FHV, SINV, and DCV, but inhibited following other types of infection. Clearly, the regulation and function of this molecule
deserves further investigation. The genes CG4680, Eip75B, Sp7,
and CG10916 are induced both by the viruses and by the intracellular bacterium Listeria (33), suggesting that they may participate in the defense against intracellular intruders (Supplemental
Table III).
A second comment is that the majority of upregulated genes are
induced by only one or two of the viruses, revealing virus-specific
responses. Of interest, 84% of the genes upregulated by SINV
are also induced by FHV, pointing to a strong similarity between
the responses to the two viruses. FHV induced a higher number of
genes than did Sindbis virus, and only 34% of the genes induced by
FHV are also induced by SINV (Fig. 5A). It is intriguing, though,
that many of the genes induced solely by FHV, but not by SINV,
are members of the same gene families as the genes coinduced by
both FHV and SINV. This peculiarity underlines the basic similarities between the transcriptional response to the two viruses. In
addition, several genes associated with cell death are induced by
FHV, but not SINV, which may reflect the higher virulence of
FHV (Fig. 5B, Supplemental Tables II, III). Only 22% and 16% of
the genes induced by SINV and FHV, respectively, are also induced by DCV, indicating that DCV, on one hand, and FHV and
SINV, on the other hand, trigger different inducible responses
(Fig. 5A). We did not detect in our microarrays expression of the
genes encoding the unpaired (Upd) cytokines, which activate the
JAK-STAT pathway in Drosophila. However, quantitative RT-PCR
analysis revealed that upd2 and upd3, but not upd, are induced or
upregulated following DCV and CrPV infection (Fig. 3C).
Virus-specific pattern of gene induction
To further characterize the transcriptional response triggered by
different viruses, wild-type flies were injected with DCV, CrPV,
FHV, SINV, VSV, DXV, and IIV-6, and gene induction was
measured at 6 h postinfection and 1, 2, 3, and 4 dpi. Gene expression was monitored by quantitative RT-PCR, which provides
a more accurate quantification of gene expression than does hybridization to short oligonucleotide probes on microarrays (34). We
monitored expression of the DCV-induced gene vir-1 (11) and of
TotM, which, according to the microarrays, is induced by FHV and
SINV infection. We confirmed the induction of vir-1 by DCV and
FHV (11) and detected a milder but significant induction of this
gene by CrPV infection. By contrast, no induction of vir-1 by
SINV, VSV, DXV, and IIV-6 was observed (Fig. 5C). For TotM,
Downloaded from http://www.jimmunol.org/ by guest on June 18, 2017
FIGURE 5. Microarray analysis of gene induction following infection by DCV, FHV, or SINV. (A) Venn diagram showing the number of upregulated
genes (by a factor of at least 2) following infection by the three viruses. The total number of genes regulated by each virus is indicated in parentheses. (B)
FHV and SINV induce members of the same gene families, but FHV triggers a stronger response. The numbers of genes belonging to seven gene ontology
functional categories induced by both FHV and SINV or by FHV only are shown. (C) Expression of vir-1 and TotM by quantitative PCR normalized for the
expression of the housekeeping gene RpL32. Groups of 10 wild-type (OR) flies were injected with Tris buffer or the viruses DCV, CrPV, FHV, SINV, VSV,
DXV, or IIV-6 or pricked with a needle dipped in a concentrated pellet of the Gram-positive bacterium M. luteus and the Gram-negative bacterium E. coli.
RNA was extracted at 6 h, 1 d, 2 d, 3 d, and 4 d after challenge. The data represent the mean and SEs of at least two independent experiments. The p values
were calculated for each time point individually versus the Tris-injected control. *p , 0.05, **p , 0.01, ***p , 0.001.
VIRUS-SPECIFIC INDUCIBLE RESPONSE IN DROSOPHILA
we confirmed the induction by FHV at different time points. In
addition, we observed that TotM expression was significantly induced by DCV at late time points of infection (4 dpi). We note that
induction of TotM by SINV, VSV, and DXV was 10–20 times
stronger than the induction by FHV (Fig. 5C). The DNA virus IIV6 did not induce TotM at any measured time point. Interestingly,
we observed different profiles for vir-1 and TotM induction after
viral challenge. Overall, the viruses that kill wild-type flies rapidly
(within 10 d), such as DCV, CrPV, and FHV, were potent inducers
of vir-1, whereas less pathogenic viruses, such as SINV, VSV, and
DXV, did not induce vir-1. The opposite trend was observed for
TotM, which was most potently induced by SINV, VSV, and DXV.
The different pattern of induction of vir-1 and TotM suggests that
the two genes may be regulated differently, even though both were
previously shown to be regulated by the JAK-STAT pathway (11,
17). Indeed, the MAP3K MEK kinase 1 (MEKK1) and the Imd
pathways are also known to contribute to the induction of TotM
induction in some contexts (17, 35).
Some antimicrobial peptide genes were also upregulated according to the microarrays, suggesting an overlap between antiviral
immunity and antibacterial–antifungal defenses. We observed an
enrichment for genes regulated by the Toll pathway [e.g., the
cytokine Spaetzle (Spz) and the antifungal peptides Drosomycine
(Drs) and Metchnikowine (Mtk)] in the DCV-specific set of genes
(Supplemental Table II). We also noted an enrichment of Imd
pathway–regulated genes, such as the antibacterial peptides AttacinA and -C, Diptericin-B, and the transcription factor Relish, in
the genes upregulated by both DCV and FHV. However, when expression of diptericin and drosomycin—two markers of activation
of the Imd and Toll pathways, respectively—was monitored by
quantitative RT-PCR, none of the viruses triggered an induction
comparable to that of bacterial and fungal infections, although the
wounding associated with the injection procedure clearly led to
some expression of the genes (Supplemental Fig. 1).
cells, as demonstrated by the critical role played by the dsRNA
receptor TLR3 in the sensing of herpesvirus infection in mammals
(39, 40). Our data are consistent with a model whereby dsRNA
generated from convergent transcription of the IIV-6 genome is
processed by Dcr-2 and triggers RNAi. Thus, we conclude that
RNAi provides an efficient and highly specific RNA-based defense against many types of viruses in Drosophila and probably
other insects. This conclusion parallels the situation described
in plants. The vertebrates, which largely rely on the induction of
IFNs to counter viral infections, appear to be the exception among
multicellular organisms (1). Of interest, however, the DExD/H
box helicase domains found in Dcr enzymes and RIG-I–like
receptors, which sense the presence of viral RNAs in cells infected
by RNA and DNA viruses, are phylogenetically related (10). This
finding suggests that an essential domain of a core molecule from
the ancestral antiviral response, RNA silencing, was at some point
recruited to sense viral RNAs in vertebrates and to subsequently
activate a signaling pathway leading to production of IFNs.
Discussion
We have investigated the involvement of RNAi and the evolutionarily conserved JAK-STAT signaling pathway in the resistance
to a panel of seven viruses representing several important families,
including the arboviruses SINV and VSV. Our data provide a
contrasting picture: on the one hand, a broad antiviral immunity
based on RNAi contributing to the defense against both RNA and
DNA viruses, and on the other hand, a virus-specific transcriptional
response involving the JAK-STAT pathway but playing a critical
role only in the case of Dicistroviridae infection.
RNAi protects against a DNA virus infection
The present study extends work from several groups, including our
own, showing that flies mutant for the siRNA pathway are more
sensitive than wild-type flies to a large panel of RNA viruses, and
reveals that Dcr-2 is also required for the control of the DNA virus
IIV-6. We note, however, that the increase of viral titer in siRNA
pathway–mutant flies is not as strong as in the case of some RNA
viruses [e.g., VSV (25)]. This finding could reflect either the expression of a viral suppressor of RNAi by IIV-6 or the fact that
only a portion of the viral genome is targeted by siRNAs. Indeed,
this virus encodes an RNaseIII enzyme, which could cleave
siRNA duplexes, as previously reported in plants infected by the
sweet potato chlorotic stunt virus (36). The involvement of Dcrmediated immune responses against DNA virus infections was
previously noted in plants, in which secondary structures in the
transcribed viral RNAs, or dsRNAs formed from overlapping bidirectional transcripts, can be processed into siRNAs (37, 38).
Production of dsRNA from DNA viruses also occurs in animal
Virus-specific induced gene expression in Drosophila
Microarrays are powerful tools to monitor the global transcriptome
of infected cells and compare the response to different infections.
Despite its limitations for accurate measurements of the magnitude
of expression changes, this technology provides useful information
on changes in gene expression (34). In this article, using wholegenome Affymetrix microarrays to analyze the transcriptome of
flies infected by DCV, FHV, or SINV, we report the existence of virusspecific responses to infection. These results are in keeping with
a previous study pointing to autophagy as an antiviral defense
mechanism against VSV, but not DCV, infection (14). The three
viruses we used belong to different families and present different
characteristics that make them valuable for the current study. For
example, 1) DCV and FHV replicate rapidly and kill Drosophila
upon injection, whereas SINV does not at the dose used (11, 24); 2)
DCV is a natural pathogen of Drosophila, whereas FHV and SINV
have not been found in wild Drosophila populations (41); 3) FHV
and DCV possess, respectively, a strong and moderate viral suppressor of RNAi, whereas SINV presumably does not (28, 42, 43).
The three viruses also have different tissue tropism and may be
associated with tissue-specific modifications in the physiology of
the infected host. For example, FHV was recently shown to be a
cardiotropic virus, affected by potassium channels regulating heart
function (44), whereas DCV infection causes intestinal obstruction
(S. Chtarbanova and J.-L. Imler, manuscript in preparation).
Comparison of the transcriptomes of the flies infected by the
three viruses revealed more similarities between FHV and SINV
than between each of these and DCV. This may reflect the coevolution of DCV with its host, and the fact that this virus may have
learned to ward off the antiviral arsenal of its host. Indeed, DCV
induces fewer genes than does FHV, even though the two viruses
replicate with similar kinetics and lead to the rapid death of the
flies. The genes induced by FHV and SINV encode chaperonins
(Tcp or Hsp), glutathione transferases, cytochrome P450s, stress
markers (Tot family), thioester-containing proteins, and cytoskeletal regulators, suggesting an involvement of oxidative stress
and phagocytosis in the response to these viruses. The two viruses
also upregulate the gene egghead (egh), which encodes a molecule
involved in the uptake of dsRNA and antiviral immunity (27).
Despite the large overlap between the genes upregulated by FHV
and SINV, the former induce a more intense transcriptional response than the latter. This observation may reflect the more aggressive replication of FHV in Drosophila. Indeed, the genes
specifically induced by FHV include not only additional members
of the families mentioned above (Hsp, Tcp, Gst, cytP450, thioester-
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656
The Journal of Immunology
as flies mutant for either RNAi or the inducible JAK-STAT
pathway succumb to infection 2–3 d before the controls, with an
∼10-fold increase in viral titer.
Interestingly, even though hop mutant flies appear to be specifically sensitive to Dicistroviridae, other viruses activate the
JAK-STAT pathway. Indeed, we observed a slight increase in the
lethality of hop mutant flies postinfection with DXV and IIV-6.
In Aedes mosquitoes, the JAK/STAT pathway was also shown to
activate a defense against Dengue, a member of the Flaviviridae
family (54). We also note that the JAK-STAT pathway–regulated
gene vir-1 (11) is induced by DCV and CrPV, but also FHV, even
though hop mutant flies resist FHV infection much as do wild-type
flies. One hypothesis to explain this apparent paradox is that some
genes may be induced in a JAK-STAT–independent manner in the
context of viral infections. For example, the gene TotM, which is
induced by several viruses normally resisted by hop mutant flies,
can be induced by the MEKK1 pathway, in addition to the JAKSTAT pathway (35). Indeed, we observed that TotM remains fully
induced by FHV and SINV in hop mutant flies (C. Dostert and
J.-L. Imler, unpublished observations). However, this hypothesis
cannot account for the induction of vir-1 by FHV, because it is
strongly reduced in hop mutant flies (C. Dostert and J.-L. Imler,
unpublished observations). This finding suggests that some aspects
of the JAK-STAT–induced response may be redundant of other
defenses for FHV, but not for DCV. The fact that FHV triggers
a stronger transcriptional response than does DCV (Fig. 5) is
consistent with this hypothesis.
A key question pertains to the nature of the receptor detecting
Dicistroviridae infection and triggering the JAK-STAT–dependent
inducible response. Our data point to the induction of a specific
subset of genes, including the JAK-STAT–regulated gene vir-1
(11), by fast-killing viruses such as DCV and CrPV, but also FHV,
which replicate rapidly to high titers upon injection in flies. Of
note, vir-1 induction is not affected in flies expressing the dsRNAbinding protein B2, or in Dcr-2 mutant flies, indicating that this
gene is not induced following sensing of dsRNA (10). This finding
suggests that sensing tissue damage and/or cell death could contribute to this inducible response, a hypothesis corroborated by the
association of the JAK-STAT pathway with the cellular response
to a variety of stresses (17, 55–57).
In conclusion, our data confirm that, beyond RNAi, an inducible
response contributes to the control of some viral infections in
Drosophila. However, this response is complex, and great care
should be exercised before generalizing the results obtained with
one single virus species. This unexpected complexity probably
reflects the intricate association of viruses with their host cells in
different tissues, their different strategies of replication or protein
expression, or their acquisition of suppressors of host defense.
Acknowledgments
Dicistroviridae-specific contribution of the JAK-STAT pathway
to antiviral immunity
An unexpected finding reported in this article is that hop mutant
flies have a clear phenotype for DCV and CrPV, but not for the
other viruses tested. This observation indicates that the JAK-STAT
pathway, in addition to RNAi, participates in host defense against
members of the Dicistroviridae family. DCV infection leads to
induction of the genes encoding the cytokines Upd2 and Upd3,
which may subsequently activate the JAK-STAT pathway in noninfected cells, triggering an antiviral program of gene expression.
Altogether, our results highlight that the contribution of the inducible response to the control of DCV is similar to that of RNAi,
We thank Estelle Santiago and Miriam Yamba for excellent technical assistance, Phil Irving for help with the microarray experiments, Anette
Schneeman (The Scripps Research Institute, La Jolla, CA) for providing
DXV and CrPV, Trevor Williams (Veracruz, Mexico) for providing IIV-6,
and Stéphanie Blandin and Dominique Ferrandon for critical reading of
the manuscript and helpful suggestions. The microarray analysis and the
deep sequencing were performed at the Plateforme Biopuces et Séquençage, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Strasbourg, France.
Disclosures
The authors have no financial conflicts of interest.
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containing proteins), supporting the idea of a more intense response,
but also genes associated with cell death. In addition, FHV upregulates several molecules previously connected to innate immunity
in Drosophila, such as Hel89B (45), POSH (46), or MEKK1 (35),
or molecules that may downmodulate the strong response to virus
infection (e.g., the genes CG9311 and Pez, encoding tyrosine
phosphatases). Finally, we note that FHV induced eight genes
encoding factors with RNA binding domains, including four
DExD/H box helicases, which may participate in the sensing and
neutralization of viral nucleic acids. This specificity may reflect
a response of the host to counter the effect of the strong suppressor
of RNAi B2, a dsRNA-binding protein (47).
An intriguing aspect of the transcriptome of virus-infected flies
is the upregulation of genes regulated by the Toll and Imd pathways.
We observed an enrichment of Toll pathway target genes induced
in flies infected by DCV, but not FHV or SINV, suggesting that
DCV infection triggers this pathway. Among the genes induced by
DCV, but not by the two other viruses, we also note the presence of
Ect4, which encodes a TIR domain cytoplasmic molecule. The
mammalian ortholog of this gene, SARM, was proposed to participate as a negative regulator of TLR signaling in some antiviral
defenses (48). Two other genes regulated by DCV and possibly
establishing a connection between RNA silencing and the inducible
response are worth mentioning: headcase was identified in a screen
as a regulator of the siRNA pathway (49), whereas CG9925 encodes a protein with a Tudor domain, a characteristic of several
components of the Piwi-interacting RNA pathway (50).
Unlike the Toll-regulated genes, several genes regulated by Imd
were induced in flies infected by DCV or FHV, although not by
SINV. The Toll and Imd pathways play a well-characterized role
in the regulation of bacterial and fungal infections, through the
regulation of genes encoding antimicrobial peptides. These genes
are also upregulated by viral infection, although not significantly,
compared with buffer injection. This low level of induction most
likely explains our inability to detect antimicrobial peptides in
the hemolymph of DCV-infected flies (51). Although not formally
establishing that the Toll and Imd pathways participate in the
antiviral response, these results certainly do not rule out such a
role (7, 9, 15). Alternatively, induction of the antimicrobial genes
may involve the transcription factor FOXO, a known regulator of
stress resistance, and may occur independently of the Toll and Imd
pathways (52). Whatever the mechanism of induction, the biological significance of this weak induction of molecules normally
active in the micromolar range is unclear. One possibility is that
the Drosophila antimicrobial peptides carry additional functions
that do not require high-level expression. For example, some
mammalian b-defensins play a dual role in innate immunity and,
in addition to their antibacterial properties, interact with chemokine receptors with affinities in the nanomolar range, thus mediating chemoattraction of phagocytic cells (53).
657
658
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Nat. Immunol. 9: 1425–1432.
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J. A. Hoffmann, and J. L. Imler. 2005. The Jak-STAT signaling pathway is required but not sufficient for the antiviral response of drosophila. Nat. Immunol. 6:
946–953.
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Drosophila C virus. J. Gen. Virol. 89: 1497–1501.
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27. Saleh, M. C., M. Tassetto, R. P. van Rij, B. Goic, V. Gausson, B. Berry,
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28. van Rij, R. P., M. C. Saleh, B. Berry, C. Foo, A. Houk, C. Antoniewski, and
R. Andino. 2006. The RNA silencing endonuclease Argonaute 2 mediates
specific antiviral immunity in Drosophila melanogaster. Genes Dev. 20: 2985–
2995.
VIRUS-SPECIFIC INDUCIBLE RESPONSE IN DROSOPHILA
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Supplemental Table II: List of genes induced by DCV, FHV and SINV
DCV specific (82 genes)
Probe number
Gene name
143500_at
rho
Synomymus
DCV *
12.5
signal transduction
Putative function
153603_at
CG10927
4.9
Unknown
151144_at
CG13553
4.2
Key words
Cytidine/deoxycytidylate deaminase zinc-binding region
152245_at
CG13335
151473_i_at
syt
synaptotagmin p65
3.4
143125_at
dl
dorsal
3.3
DNA binding
NF-kappa-B/Rel/dorsal
150552_at
CG9996
3.1
Unknown
Glycosyl transferase family 8
143770_at
Mtk
Metchnikowin
3.0
Antifungal peptide activity
141691_at
spz
spatzle
141444_at
CG2471
141366_at
CG10764
154443_at
CG1667
151661_s_at
Nrt
141718_at
D12
147495_at
CG10081
143869_at
vri
3.6
serine protease
Neurotactin
enzyme
3.0
Toll binding
3.0
actin binding
Leucine-rich repeat
3.0
serine-type endopeptidase activity
Peptidase trypsin-like serine and cysteine proteases
2.9
unknown
2.8
cell adhesion
Esterase/lipase/thioesterase
2.8
transcription factor
YEATS family
2.8
Unknown
Peptidase M28
vrille
2.8
RNA polymerase II transcription factor activity
Basic-leucine zipper (bZIP) transcription factor
Calpain-B
2.7
calpain activity
Peptidase C2 calpain
Unknown
C-type lectin
AICARFT/IMPCHase bienzyme, Methylglyoxal synthase-like
domain
151648_at
CalpB
152979_at
CG31531
2.7
152405_at
CG6014
2.7
147951_at
CG13905
2.7
154983_at
CG15634
2.7
153761_at
CG11089
2.6
Unknown
141974_at
CG4573
2.6
tyrosine--tRNA ligase; enzyme
153004_at
CG8603
2.6
motor
153432_at
Thor
2.6
Eukaryotic initiation factor 4E binding
Eukaryotic translation initiation factor 4E binding
142200_at
CG32477
151361_at
RhoGAP18B
Rho GTPase activation protein
148533_at
ninjurin A
Thor
2.5
2.5
GTPase activator activity
ninjurin
2.5
cell adhesion
myosin I
2.5
Actin binding
Myosin head (motor domain), IQ calmodulin-binding region
2.5
v-SNARE activity
Vesicle transport v-SNARE
2.5
netrin receptor activity
142429_at
CG14089
142097_at
Myosin 31DF
2.5
154415_at
CG3279
143665_at
fra
153174_at
CG9925
151557_i_at
CG32737
2.5
144197_at
rdgB
2.5
147151_at
CG18279
2.4
frazzled
Collagen triple helix repeat
2.5
headcase
phosphatidylinositol transporter activity
Phosphatidylinositol transfer protein
155065_at
hdc
151217_at
U snoRNA host gene 1
2.3
151871_at
CG30494
2.3
144505_at
CG32767
2.3
Nucleic acid binding
Zn-finger C2H2 type, Cytochrome c heme-binding site,
Immunoglobulin/MHC
152889_at
dei
2.3
transcription factor activity
Basic helix-loop-helix dimerization domain bHLH
153798_at
CG10103
2.3
transcription factor
Eukaryotic protein of unknown function DUF292, Pancreatic
ribonuclease
154436_at
rtet
2.3
sugar porter activity
General substrate transporter, Sugar transporter
superfamily
147575_at
CG18413
142250_at
norpA
153304_at
bip1
delilah
tetracycline resistance
2.3
Fibronectin type III, Immunoglobulin-like, Peptidase M14
carboxypeptidase A
Maternal tudor protein, Zn-finger MYND type
signal transduction
ligand binding or carrier
2.3
1-phosphatidylinositol-45biphosphate phosphodiesterase
2.3
1-phosphatidylinositol-45-biphosphate
phosphodiesterase ; EC:3.1.4.11
2.2
145526_at
CG4213
2.2
motor
148109_at
CG12014
2.2
iduronate-2-sulfatase activity
Sulfatase
153934_at
CG12190
2.2
transcription factor
Zn-finger Ran-binding
153814_at
CG17612
143293_at
pbl
143726_at
Dhc93AB
155084_at
148881_at
2.2
nucleic acid binding
Zn-finger C2H2 type
pebble
2.2
guanyl-nucleotide exchange factor activity
Rhodopsin-like GPCR superfamily
Dynein heavy chain at 93AB
2.2
Dynein_heavy;microtubule motor activity
Dynein heavy chain, Peptidase eukaryotic cysteine
peptidase active site, AAA ATPase
CG17260
2.2
nucleic acid binding
Zn-finger RING
CG13059
2.2
154573_at
CG8149
2.2
SAP;DNA binding;4
DNA-binding SAP
146703_at
CG30158
2.2
GTP binding
Ras GTPase superfamily
153520_at
5-HT7
2.2
G-protein coupled serotonin receptor activity
Rhodopsin-like GPCR superfamily
154930_at
CG3165
154682_at
Krn
146238_at
CG32955
Serotonin receptor 7
3'-5' exonuclease
2.1
enzyme
Keren
2.1
EGF receptor binding
2.1
kinesin;motor activity
EGF-like domain
143507_at
mys
integrin-beta-subunit
2.1
cell adhesion receptor
143605_at
Drs
Drosomycin
2.1
antifungal peptide activity
Gamma thionin, Knottin
151022_at
CG33188
2.1
protein binding
Zn-finger AN1-like
147490_at
Rgk1
2.1
RAS small monomeric GTPase activity
Ras GTPase superfamily
148775_at
CG10006
GTP binding protein-like
2.1
Zip;metal ion transporter activity
Zinc transporter ZIP
152824_at
Ect4
2.1
153034_at
CG32183
2.1
growth factor activity
Thrombospondin type I, von Willebrand factor type C,
Cystine knot C-terminal
transcription factor
Homeodomain-like, ARID (AT-rich interaction domain)
protein
Sterile alpha motif SAM, ARM repeat fold, TIR
144380_at
CG14770
2.1
151685_r_at
CG32813
2.1
148681_at
CG12520
2.1
154808_at
CG4953
2.1
151834_at
CG32130
2.0
BAG;protein binding
141962_at
CG5787
2.0
transcription factor; DNA binding
2.0
calcium potassium:sodium antiporter activity
151600_at
Nckx30C
148381_r_at
CG13674
sodium/calcium exchanger
141379_at
Ag5r
148130_at
ida
143472_at
Ten-m
143221_at
Gliotactin
150044_at
CG18213
2.0
146411_at
CG15145
2.0
154066_at
wus
Protein of unknown function DUF974, Actinoxanthin-like
Peptidase S1 chymotrypsin, Apoptosis regulator Bcl-2
protein BAG
Sodium/calcium exchanger membrane region, K+dependent Na+/Ca+ exchanger related-protein
2.0
Antigen 5-related
2.0
imaginal discs arrested
2.0
Tenascin major
2.0
cell adhesion
2.0
serine esterase
gliotactin
wurst
2.0
Allergen V5/Tpx-1 related
TPR-like
EGF-like domain
Zn-finger MYND type
chaperone
Heat shock protein DnaJ N-terminal
FHV specific (279 genes)
Probe number
Gene name
143470_at
LysX
141566_at
CG10805
152381_at
Pcaf
152230_at
CG6778
155116_at
POSH
152907_at
18w
152730_at
149122_at
Synomymus
Lysozyme X
FHV *
6.9
Putative function
lysozyme activity, cell death
IPBinding-protein-dependent transport systems inner
membrane component, Aldo/keto reductase, HEAT
6.0
histone acetyltransferase
Key words
Glycoside hydrolase family 22
5.3
histone acetyltransferase activity
4.5
glycine--tRNA ligase
Bromodomain, GCN5-related N-acetyltransferase
Plenty of SH3s
4.3
receptor signaling complex scaffold activity
Zn-finger RING, Neutrophil cytosol factor 2
18 wheeler
4.2
transmembrane receptor activity
Leucine-rich repeat, TIR, Interleukin-1 receptor type I/Toll
precursor
CG5205
4.2
RNA helicase activity
CG5571
4.0
144194_at
Dgp-1
3.8
147210_at
CG12868
142986_at
CG9311
143828_at
145304_at
154332_at
146014_at
143953_at
Cas
153828_at
Sec63 domain, DEAD/DEAH box helicase, AAA ATPase
Rhodopsin-like GPCR superfamily
GTP binding, hydrolase activity acting on acid
anhydrides in phosphorus-containing
anhydrides
Protein synthesis factor GTP-binding, Elongation factor Tu
domain 2
Tyrosine specific protein phosphatase, Rhodopsin-like
GPCR superfamily
3.8
protein phosphatase-like
3.7
protein-tyrosine-phosphatase activity
Apc
APC-like
3.6
beta-catenin binding, microtubule binding
Armadillo, APC cysteine-rich
psh
persephone
3.6
serine-type endopeptidase activity
Peptidase S1 chymotrypsin
CG12876
ALix
3.6
signal transducer activity, cell death
BRO1, Rho GTPase activation protein
CG8353
cytidine deaminase-like
3.6
cytidine deaminase activity,
dCMP_cyt_deam;zinc ion binding
Cytidine/deoxycytidylate deaminase zinc-binding region
CAS/CSE1 segregation protein
3.6
importin-alpha export receptor activity, cell
death, phagocytosis
CAS/CSE C-terminal, Importin-beta N-terminal
CG15828
retinoid binding/fatty acid binding
protein-like glycoprotein
3.5
lipid binding, Vitellogenin_N;lipid transporter
activity
von Willebrand factor type D, Lipid transport protein Nterminal, Protein of unknown function DUF1081
141776_at
CG12428
peroxisomal carnitine Ooctanoyltransferase
3.4
carnitine O-octanoyltransferase activity
Acyltransferase ChoActase/COT/CPT
145889_at
Pez
protein phosphatase-like
3.4
protein-tyrosine-phosphatase activity,
cytoskeletal protein binding
Tyrosine specific protein phosphatase and dual specificity
protein phosphatase
Glutathione S-transferase
149757_at
GstD3
Glutathione S transferase D3
3.4
glutathione transferase activity
143031_at
CG7627
ATP-binding cassette transporter
3.4
ATP-binding cassette (ABC) transporter activity ABC transporter, AAA ATPase
145025_at
CG12177
purine nucleosidase
3.3
enzyme
Inosine/uridine-preferring nucleoside hydrolase
149173_at
CG7632
3.3
enzyme
Annexin, Esterase/lipase/thioesterase
150261_at
CG4335
3.3
gamma-butyrobetaine dioxygenase activity
Gamma-butyrobetaine hydroxylase
Serine/threonine protein kinase
Mekk1
heat shock construct of Inoue
3.3
receptor signaling protein serine/threonine
kinase activity, MAP kinase kinase kinase
activity
149754_at
GstD10
Glutathione S transferase D10
3.3
glutathione transferase activity
Glutathione S-transferase
152949_at
Cyp6d4
cytochrome P450 CYP6D4
3.2
cytochrome P450 activity
Cytochrome P450, E-class P450 group I
152798_at
Cyp12c1
cytochrome P450 CYP12C1
3.2
cytochrome P450 activity
Cytochrome P450, E-class P450 group I
153844_at
Rs1
3.2
ATP dependent RNA helicase activity,
DEAD;ATP binding
DEAD/DEAH box helicase, ATP-dependent helicase DEADbox
143384_at
trx
3.2
DNA binding
Zn-finger-like PHD finger, FY-rich domain SET-related region
141756_at
trithorax
141436_at
CG3505
3.2
serine-type endopeptidase activity
Peptidase trypsin-like serine and cysteine proteases
152481_at
CG9951
3.2
motor
Protein of unknown function DUF812
141381_at
CG30069
3.2
RNA_pol_Rpb1_R;DNA binding
Short-chain dehydrogenase/reductase SDR
La autoantigen-like
3.2
Pol III transcription termination factor activity,
RNA binding, 5S rRNA primary transcript
binding
RNA-binding region RNP-1 (RNA recognition motif), RNAbinding protein Lupus Lal
3.1
glutathione transferase activity
Glutathione S-transferase
Tcp1-like
3.1
chaperone activity, phagocytosis
Chaperonin Cpn60/TCP-1, GroEL-like chaperone ATPase
143668_at
La
153930_at
CG5224
143374_at
T-cp1
153051_at
CG4841
3.1
143696_at
CG4038
3.0
141466_at
CG15099
154265_at
CG8258
151745_at
CG11033
3.0
Cyclin-like F-box, Zn-finger CXXC type, Cytochrome c hemebinding site, Leucine-rich repeat cysteine-containing
subtype, Transcription factor jumonji jmjC
152411_at
CG10286
3.0
ARM repeat fold
146573_at
CG9253
3.0
152638_at
CG3363
154861_at
asf1
152420_at
Hel89B
152946_at
Uba1
151728_at
CG18596
T-complex protein 1 theta-subunit
Gar1 protein RNA-binding region
3.0
protein kinase
Dopey N-terminal
3.0
chaperonin ATPase activity
Chaperonin Cpn60/TCP-1, GroEL-like chaperone ATPase
ATP dependent RNA helicase activity
DEAD/DEAH box helicase, Heavy metal
transport/detoxification protein
2.9
DNA binding
anti-silencing factor 1
2.9
cell cycle regulator
Anti-silencing protein ASF1-like, p53-like transcription factor
Helicase 89B
2.9
ATP dependent DNA helicase activity
SNF2 related domain, DEAD/DEAH box helicase, ARM repeat
fold
Ubiquitin activating enzyme 1
2.9
ubiquitin activating enzyme activity
UBA/THIF-type NAD/FAD binding fold, Ubiquitin-activating
enzyme repeat
2.9
SpoU_methylase;RNA binding
tRNA/rRNA methyltransferase (SpoU), ARM repeat fold
147940_at
CG13900
spliceosomal protein-like
2.9
damaged DNA binding, CPSF_A;nucleic acid
binding
CPSF A subunit C-terminal, Immunoglobulin/major
histocompatibility complex
Actin/actin-like, phagocytosis
154742_at
Arp66B
Actin-related protein 66B
2.8
actin binding, structural constituent of
cytoskeleton
141603_at
Aats-val
Valyl-tRNA synthetase
2.8
glutamate-tRNA ligase activity
Aminoacyl-tRNA synthetase class Ia, Valyl-tRNA synthetase
class Ia
147841_at
Zfrp8
Zinc finger protein RP-8
2.8
DNA binding
Zn-finger MYND type, Programmed cell death protein 2 Cterminal, 4Fe-4S ferredoxin iron-sulfur binding domain
151838_at
CG2789
154028_at
CG9200
154859_at
CG17904
152023_at
TepIII
146827_at
CG8738
150418_at
sav
153299_at
CG12292
152623_at
Est2
154156_at
CG5033
148662_at
CG10638
154452_at
c11.1
154670_at
CG3476
143144_at
eg
154451_at
CG2691
143782_at
Cyp4p1
153336_at
Aats-asp
141481_at
CG32702
149333_at
PEK
145970_at
TepII
142141_at
CG4723
141207_at
CG4453
peripheral-type benzodiazepine
receptor
Thiolester containing protein III
salvador
2.8
benzodiazepine receptor activity
TspO/MBR-related protein
2.8
myb_DNA-binding;DNA binding
Myb DNA-binding domain, Homeodomain-like
2.8
nucleotide binding
2.8
endopeptidase inhibitor activity, phagocytosis
Alpha-2-macroglobulin, Terpenoid cylases/protein
prenyltransferase alpha-alpha toroid
2.8
serine-type endopeptidase activity
Peptidase trypsin-like serine and cysteine proteases
2.8
enzyme, cell death
WW/Rsp5/WWP domain
2.8
Esterase-2
aldehyde reductase-like
Protein of unknown function DUF803
2.7
carboxylesterase activity
Carboxylesterase type B, Esterase/lipase/thioesterase
2.7
transcription factor activity
G-protein beta WD-40 repeat
2.7
aldehyde reductase activity, trans-12Aldo/keto reductase
dihydrobenzene-12-diol dehydrogenase activity
2.7
ARM repeat fold
carnitine-acylcarnitine carrier
protein
2.7
carrier activity, acyl carnitine transporter
activity
Mitochondrial substrate carrier, Mitochondrial carrier
protein, Adenine nucleotide translocator 1
eagle
2.7
transcription factor activity, ligand-dependent
nuclear receptor activity
Zn-finger C4-type steroid receptor, Vitamin D receptor,
Steroid nuclear receptor ligand-binding
2.7
ARM repeat fold
Cytochrome P450-4p1
2.7
cytochrome P450 activity
Cytochrome P450, E-class P450 group I
Aspartyl-tRNA synthetase
2.7
aspartate-tRNA ligase activity
tRNA synthetase class II (D K and N), OB-fold nucleic acid
binding domain, Aspartyl-tRNA synthetase archea/euk type
2.7
metalloendopeptidase activity
CUB, EGF-like domain, Aspartic acid and asparagine
hydroxylation site
2.7
protein kinase activity, eukaryotic elongation
factor-2 kinase activity
Serine/threonine protein kinase, Bacterial quinoprotein
2.7
antibacterial peptide activity, wide-spectrum
protease inhibitor activity, phagocytosis
Alpha-2-macroglobulin, Serpin, Terpenoid cylases/protein
prenyltransferase alpha-alpha toroid
Peptidase M13 neprilysin
type-I transmembrane ER-resident
serine/threonine kinase-like
Thiolester containing protein II
nucleoporin
moleskin
2.7
endothelin-converting enzyme activity,
Peptidase_M13_N;metallopeptidase activity
2.7
endopeptidase
2.6
protein carrier activity, RAN protein binding,
protein transporter activity
151799_at
msk
144700_at
CG15347
2.6
Protein of unknown function DUF753
146788_at
CG11210
2.6
Protein of unknown function DUF221
149968_at
CG5399
2.6
154083_at
Smg5
2.6
152479_at
CG6734
2.6
sphingomyelin phosphodiesterase activator
activity
Beige/BEACH domain, G-protein beta WD-40 repeat,
Immunoglobulin/MHC
142720_at
CG11660
142550_at
l(3)mbt
145098_at
Lsd-2
Importin-beta N-terminal, ARM repeat fold
Nucleotide binding protein PINc
2.6
protein kinase activity
Protein of unknown function RIO1, Tyrosine protein kinase
lethal (3) malignant brain tumor
2.6
transcription factor activity
Sterile alpha motif (SAM)/Pointed domain, Mbt repeat
Lipid storage droplet-2
2.6
Perilipin
154565_at
CG12007
2.6
RAB-protein geranylgeranyltransferase activity,
Protein prenyltransferase
PPTA;protein prenyltransferase activity
145074_at
CG5321
2.6
gamma-butyrobetaine dioxygenase activity
Gamma-butyrobetaine hydroxylase
145681_at
CG31694
interferon-like
2.6
signal transduction
Interferon-related protein conserved region, Ankyrin, ARM
repeat fold
pastrel
2.6
Thiolester containing protein IV
2.6
antibacterial peptide activity, endopeptidase
inhibitor activity
Alpha-2-macroglobulin, HPr serine phosphorylation site
153088_at
pst
146503_at
TepIV
154692_at
CG18240
2.6
141564_at
CG2017
2.6
GTP binding, GTP_EFTU;translation elongation Protein synthesis factor GTP-binding, Elongation factor Tu
factor activity
domain 2
146823_at
Cirl
2.6
latrotoxin receptor activity, Gal_Lectin;sugar
binding
147690_at
Ugt58Fa
2.6
glucuronosyltransferase activity,
UDPGT;transferase activity transferring hexosyl UDP-glucoronosyl/UDP-glucosyl transferase
groups
153330_at
Nop56
2.6
latrophilin receptor-like
2.5
Cytochrome P450, E-class P450 group I
D-galactoside/L-rhamnose binding SUEL lectin domain, Gprotein coupled receptor family 2 (secretin-like)
Pre-mRNA processing ribonucleoprotein binding region,
Nucleic acid-binding OB-fold
154194_at
CG8939
142182_s_at
Ppn
Papilin
2.5
rRNA methyltransferase activity
Ribosomal RNA methyltransferase RrmJ/FtsJ
143690_at
Fdxh
Ferredoxin
2.5
electron carrier activity, fer2;electron
transporter activity
Ferredoxin, Adrenodoxin, Cytochrome c heme-binding site
Zinc-containing alcohol dehydrogenase superfamily,
Thioesterase, Phosphopantetheine-binding domain, Acyl
transferase domain
Fibronectin type III, Ribosomal protein S2
Pancreatic trypsin inhibitor (Kunitz), Immunoglobulin-like,
Thrombospondin type I, EGF-like domain
151887_at
v(2)k05816
2.5
[acyl-carrier protein] S-acetyltransferase
activity, enoyl-[acyl-carrier protein] reductase
(NADPH B-specific) activity
154537_at
CG31738
2.5
cell adhesion
144074_at
CG4025
2.5
148524_at
CG14152
2.5
149907_r_at
CG8066
154829_at
CG7728
cystatin-like
149345_at
CG12171
152727_at
CG7224
153358_at
Nmd3
153152_at
CG11920
154616_at
CG31633
154916_at
CG7417
145612_at
CG7291
141658_at
CG5853
ATP-binding cassette transporter
142836_at
CG32041
heat shock protein 22 kD
146844_at
CG30349
143606_at
Cyp18a1
155054_at
2.5
cysteine protease inhibitor activity
2.5
alcohol dehydrogenase-like
2.5
Nonsense-mediated mRNA 3
2.5
Sarcocystatin, Cysteine protease inhibitor
Protein of unknown function DUF663
oxidoreductase activity acting on CH-OH group Short-chain dehydrogenase/reductase SDR, Glucose/ribitol
of donors, adh_short;oxidoreductase activity
dehydrogenase
2.5
RNA binding, protein binding
2.5
alpha-tocopherol transfer proteinlike
Brix domain
2.5
ligand binding or carrier
Cyclin-like F-box, Four-helical cytokine
2.5
transcription factor activity
Ubiquitin system component Cue, Zn-finger Ran-binding
E1 protein and Def2/Der2 allergen
2.5
signal transduction
2.5
ATP-binding cassette (ABC) transporter activity Phagocytosis, Phosphopantetheine attachment site
2.5
heat shock protein
2.5
Cytochrome P450-18a1
NMD3 family, Cytochrome c heme-binding site
G-protein beta WD-40 repeat
2.5
cytochrome P450 activity
Cytochrome P450, E-class P450 group I
CG18143
2.5
guanine deaminase activity,
Amidohydro_1;hydrolase activity
Amidohydrolase, Calcium-binding EF-hand
152398_at
CG10631
2.5
nucleic acid binding
Zn-finger C2H2 type, Zinc-finger protein of unknown
function DM3
154928_at
wdn
2.4
RNA polymerase II transcription factor activity
Zn-finger C2H2 type
wings down
145309_at
CG6335
histidine--tRNA ligase
2.4
enzyme
142167_at
CG17259
serine--tRNA ligase-like
2.4
serine-tRNA ligase activity
145230_at
CG5010
2.4
142244_at
CG9305
2.4
153052_at
CG10420
2.4
154647_at
CG8877
143040_at
CG4797
142263_at
CG32836
151511_r_at
dom
153862_at
l(3)07882
153458_at
Rep
141309_at
CG7611
152222_at
CG4445
sugar transporter-like
domino
tRNA synthetases class-II (G H P and S), Aminoacyl-transfer
RNA synthetase class II
CHCH
transcription factor activity, myb_DNAbinding;DNA binding
Myb DNA-binding domain
ARM repeat fold
2.4
pre-mRNA splicing factor activity
Peptidase M67 Mov34
2.4
glucose transporter activity,
sugar_tr;transporter activity
General substrate transporter, Sugar transporter
superfamily
2.4
ligand binding or carrier
2.4
helicase activity, DNA dependent ATPase
activity, general RNA polymerase II
transcription factor activity
2.4
SNF2 related domain, DNA topoisomerase II, ATP-dependent
DNA ligase
Nop14-like protein
2.4
RAB-protein geranylgeranyltransferase activity,
Rab GTPase activator
Rab escort protein activity
2.4
chaperone
polypeptide Nacetylgalactosaminyltransferase
2.4
polypeptide N-acetylgalactosaminyltransferase
Glycosyl transferase family 2, Ricin B lectin domain
activity
Rab escort protein
146718_at
Tsp42Er
Tetraspanin 42Er
2.4
143976_at
Pp4-19C
Protein phosphatase 19C
2.4
protein serine/threonine phosphatase activity,
CTD phosphatase activity,
Metallophos;hydrolase activity
cricklet
2.3
carboxylesterase activity
G-protein beta WD-40 repeat, Lissencephaly type-1-like
homology motif
CD9/CD37/CD63 antigen, Tetraspanin
Serine/threonine-specific protein phosphatase and bis(5nucleosyl)-tetraphosphatase
Carboxylesterase type B, TONB Box N terminus,
Esterase/lipase/thioesterase
154994_at
clt
145265_at
CG6762
2.3
154620_at
CG6181
2.3
151373_at
CG32795
2.3
147699_at
CG4250
2.3
152100_at
CG31705
2.3
153963_at
CG16971
2.3
153320_at
Smg6
2.3
RNA binding
Nucleotide binding protein PINc, TPR-like
152357_at
Rab-RP4
2.3
RAB small monomeric GTPase activity, GTP
binding, ras;small monomeric GTPase activity
Ras GTPase superfamily, ARF/SAR superfamily, Sigma-54
factor interaction domain
2.3
ATP dependent RNA helicase activity,
KH;nucleic acid binding
KH, DEAD/DEAH box helicase, ATP-dependent helicase
DEAD-box
154657_at
Rab-related protein 4
CG7878
ParB-like nuclease
motor
G-protein beta WD-40 repeat
EGF-like domain
DNA binding
143627_at
cora
coracle
2.3
cytoskeletal protein binding
Band 4.1, Ezrin/radixin/moesin ERM, FERM
153565_at
l(2)gl
lethal (2) giant larvae
2.3
myosin II binding, cell death
Lethal(2) giant larvae protein, Regulator of chromosome
condensation RCC1
144536_at
Tre1
heat shock construct of Ishimoto
2.3
taste receptor activity, G-protein coupled
receptor activity unknown ligand, cell death
Rhodopsin-like GPCR superfamily
modulo
2.3
DNA binding, RNA binding, rrm;nucleic acid
binding
RNA-binding region RNP-1 (RNA recognition motif)
refractory to sigma P
2.3
154081_at
mod
152056_at
ref(2)P
Octicosapeptide/Phox/Bem1p, Zn-finger ZZ type, Peptidase
eukaryotic cysteine peptidase, Ubiquitin-associated domain
153126_at
CG11523
2.3
145975_at
CG13795
2.3
SNF;neurotransmitter:sodium symporter
activity
152368_at
CG8913
2.3
peroxidase activity
Animal haem peroxidase, Haem peroxidase
142736_s_at
Hmgs
2.3
hydroxymethylglutaryl-CoA synthase activity
Hydroxymethylglutaryl-coenzyme A synthase
cyclin binding, NAP;DNA binding
Nucleosome assembly protein (NAP)
HMG Coenzyme A synthase
154193_at
Set
2.3
149558_at
CG16777
2.3
146490_at
CG9978
147547_at
CG18067
142395_at
CG31839
nimrodB2
2.3
151642_at
stich1
sticky ch1
2.3
galactose-specific C-type lectinlike
2.3
lectin_c;sugar binding
2.3
Sodium:neurotransmitter symporter
C-type lectin
3'5'-cyclic nucleotide phosphodiesterase
cell adhesion
Protein of unknown function DUF139, EGF-like domain,
Growth factor receptor
Basic helix-loop-helix dimerization domain bHLH
141751_at
east
144771_at
CG9691
2.3
150295_at
CG7044
2.3
145684_at
CG17224
2.3
152444_at
ken
148573_at
CG7560
151946_at
Os9
enhanced adult sensory threshold
2.3
Peptidase M14 carboxypeptidase A
ARM repeat fold
uridine phosphorylase activity,
PNP_UDP_1;catalytic activity
Purine and other phosphorylases family 1, Uridine
phosphorylase eukaryotic
ken and barbie
2.3
transcription factor activity, phagocytosis
Zn-finger C2H2 type, BTB/POZ domain, Immunoglobulin-like
methylenetetrahydrofolate
reductase
2.3
methylenetetrahydrofolate reductase (NADPH)
activity
Methylenetetrahydrofolate reductase
Olfactory-specific 9
2.3
pheromone binding
152518_at
CG8231
T-complex protein 1 zeta-subunit
2.3
chaperonin ATPase activity
152430_at
l(2)01424
NAT1
2.3
translation initiation factor activity, MIF4G;RNA eIF4-gamma/eIF5/eIF2-epsilon, Initiation factor eIF-4 gamma
binding, cell death
middle, ARM repeat fold
152280_at
CG2467
147131_at
Spt-I
2.3
Chaperonin Cpn60/TCP-1, GroEL-like chaperone ATPase
Endoglin/CD105 antigen
Serine palmitoyltransferase
subunit I
2.2
serine C-palmitoyltransferase activity
Glutathione S transferase D5
2.2
glutathione transferase activity
149759_at
GstD5
152817_at
CG1648
153515_at
Dr
144950_at
CG32654
149937_at
CG6912
142364_at
HBS1
143646_at
Pbprp1
154719_at
CG1234
146710_at
Tsp42Ef
144142_at
Kaz1
2.2
serine protease inhibitor
154940_at
CG6876
2.2
pre-mRNA splicing factor activity
146453_at
CG31753
141644_at
kis
147097_at
CG30044
143699_at
Ser6
152874_at
CG30015
Glutathione S-transferase
2.2
Drop
glutamate receptor-like
2.2
specific RNA polymerase II transcription factor
Homeobox, Homeodomain-like
activity
2.2
enzyme
2.2
translation initiation factor 3-like
2.2
translation release factor activity,
Elongation factor Tu, Translation factor, Protein synthesis
GTP_EFTU;translation elongation factor activity factor GTP-binding
Pheromone-binding proteinrelated protein 1
2.2
pheromone binding, phenylalkylamine binding, Pheromone/general odorant binding protein PBP/GOBP,
PBP_GOBP;odorant binding
Insect pheromone/odorant binding protein PhBP
CBF/Mak21 family, Peptidase eukaryotic cysteine peptidase,
ARM repeat fold
2.2
Tetraspanin 42Ef
2.2
CD9/CD37/CD63 antigen, Tetraspanin
Pre-mRNA processing ribonucleoprotein binding region
2.2
nucleic acid binding
kismet
2.2
DNA helicase activity
BRK, SNF2 related domain, Helicase, Chromo, DEAD/DEAH
box helicase
Serine protease 6
2.2
serine-type endopeptidase activity,
trypsin;trypsin activity
Peptidase trypsin-like serine and cysteine proteases
2.2
motor
2.2
144115_at
cib
144890_at
CG15220
ciboulot
2.2
actin binding
Thymosin beta-4
replication factor A 14 kD subunit
2.2
DNA binding
Nucleic acid-binding OB-fold
149419_at
CG1105
146699_at
Spn4
Serine protease inhibitor 4
2.2
serine-type endopeptidase inhibitor activity
Serpin, Endoplasmic reticulum targeting sequence
142349_at
yellow-f
yellow-f
149057_at
CG9295
cuticle protein
2.2
intramolecular isomerase activity
Major royal jelly protein
2.2
structural constituent of cuticle (sensu Insecta) Insect cuticle protein
154058_at
CG5295
2.2
Patatin;catalytic activity
142674_at
CG8282
154433_at
CG7338
151902_at
jbug
2.2
sorting nexin-like
jitterbug
Arrestin
Patatin
2.2
Phox-like
2.2
Protein of unknown function DUF663
2.2
actin binding
Filamin/ABP280 repeat, Calponin-like actin-binding, Actinbinding actinin-type
2.2
Ribosomal_L7Ae;structural constituent of
ribosome
Ribosomal protein L7Ae/L30e/S12e/Gadd45, High mobility
group-like nuclear protein
2.2
enzyme inhibitor
Ankyrin, Ribosomal protein L9 N-terminal-like
2.2
Ras GTPase activator activity
C2 domain, Pleckstrin-like, Ras GTPase-activating protein,
Rho GTPase activation protein
142579_at
NHP2
141491_at
CG8465
143456_at
Gap1
154505_at
CG9107
2.2
rrm;nucleic acid binding
RNA-binding region RNP-1 (RNA recognition motif)
154039_at
CG11738
2.2
RNA binding
KH
141457_at
Mtor
Megator
2.2
motor
141622_at
CG5841
ankyrin-like
2.1
apoptosis inhibitor
Ankyrin, Zn-finger ZZ type, Mib_herc2, Zn-finger RING
149624_at
CG3999
glycine dehydrogenase p protein
2.1
glycine dehydrogenase (decarboxylating)
activity
Glycine cleavage system P-protein
154013_at
CG1753
cystathionine beta-synthase-like
2.1
cystathionine beta-synthase activity,
PALP;lyase activity
Cysteine synthase/cystathionine beta-synthase Pphosphate-binding site, Cystathionine beta-synthase
141461_at
Alas
Aminolevulinate synthase
2.1
154735_at
CG8862
155000_at
CG7806
149590_at
CG8516
150379_at
CG6937
141762_at
Cp1
151904_at
CG31012
155134_at
MICAL-like
143738_at
Nsf2
GTPase-activating protein 1
2.1
ATP-binding cassette transporter
2.1
Aminotransferase class I and II, Aminolevulinic acid
synthase
endonuclease G activity, Endonuclease;nucleic
DNA/RNA non-specific endonuclease
acid binding
5-aminolevulinate synthase activity
ion channel
2.1
RNA binding protein-like
2.1
RNA binding, rrm;nucleic acid binding
Cysteine proteinase-1
2.1
cathepsin L activity, KRAB;nucleic acid binding, Peptidase C1A papain, KRAB box, Peptidase eukaryotic
Cell death
cysteine peptidase active site
RNA-binding region RNP-1 (RNA recognition motif)
2.1
SH3/SH2 adaptor protein activity
SH3, Neutrophil cytosol factor 2
alpha-actinin-like
2.1
actin binding
Zn-binding protein LIM, Calponin-like actin-binding
NEM-sensitive fusion protein 2
2.1
cdc48_N;ATP binding
AAA ATPase VAT, Cell division protein 48 CDC48 domain 2,
Aspartate decarboxylase-like fold
143000_at
CG5525
144243_at
Spn6
154116_at
SNF4A
149138_at
CG4858
2.1
chaperonin ATPase activity
Chaperonin Cpn60/TCP-1, GroEL-like chaperone ATPase
Serine protease inhibitor 6
2.1
serine-type endopeptidase inhibitor activity
Serpin
SNF4/AMP-activated protein
kinase gamma subunit
2.1
protein serine/threonine kinase activity,
SNF1A/AMP-activated protein kinase activity
CBS domain
2.1
nucleotide binding
Mrp family
146386_at
CG13284
estradiol 17 beta-dehydrogenaselike
2.1
steroid dehydrogenase activity,
adh_short;oxidoreductase activity
Short-chain dehydrogenase/reductase SDR, Glucose/ribitol
dehydrogenase, phagocytosis
145647_at
Cyp309a1
cytochrome P450 CYP309A1
2.1
cytochrome P450 activity
Cytochrome P450, E-class P450 group I
147354_at
CG15917
152511_at
Ela
Elastin-like
2.1
142566_at
CG2926
2.1
PHD;DNA binding
Zn-finger RING, Zn-finger-like PHD finger
2.1
serine-type endopeptidase inhibitor activity
Serpin
152581_at
Spn5
152470_at
CG10189
2.1
Serine protease inhibitor 5
Collagen triple helix repeat
2.1
152019_at
to
155057_at
CG10107
takeout
2.1
2.1
153909_at
CG2972
2.1
Odorant binding protein
cysteine-type peptidase activity
Peptidase C48 SUMO/Sentrin/Ubl1
Protein of unknown function DUF133
149260_s_at
CG10712
2.1
DNA binding
145393_at
CG32528
2.1
actin binding
145547_r_at
CG13947
2.1
152066_at
BcDNA:GH06451
asparagine--tRNA ligase-like
2.1
enzyme
144015_at
Smrter
Smr
2.1
transcription co-repressor, phagocytosis
ecdysone receptor co-repressor
141553_at
CG9139
Rabex-5, guanyl-nucleotide
exchange factor
2.1
guanyl-nucleotide exchange factor activity, zfA20;DNA binding
Zn-finger A20-like, Vacuolar sorting protein 9
149020_at
CG6843
CBF1 interacting corepressor
2.1
transcription factor binding
154133_at
CG15817
2.1
ubiquitin-specific protease activity,
UCH;cysteine-type endopeptidase activity
Peptidase C19 ubiquitin carboxyl-terminal hydrolase family
2
154252_at
LanA
2.1
structural protein
Laminin-type EGF-like domain, Galactose-binding like,
Concanavalin A-like lectin/glucanase
Laminin A
145466_at
CG32521
2.1
150514_at
CG13618
2.1
141539_at
Trx-2
152780_at
CG7967
147696_at
CG13510
142657_at
PGRP-SB1
thioredoxin-2
2.1
Calponin-like actin-binding
Odorant binding protein
thiol-disulfide exchange intermediate activity,
thiored;electron transporter activity
2.1
Thioredoxin type domain, Thioredoxin domain 2
Protein of unknown function DUF605
2.1
peptidoglycan recognition proteinlike
N-acetylmuramoyl-L-alanine amidase family 2, Animal
peptidoglycan recognition protein PGRP
2.1
peptidoglycan recognition activity
Translation initiation factor IF5
142531_at
eIF-2
Eukaryotic initiation factor 2&bgr;
2.1
tRNA binding, translation initiation factor
activity, GTP binding
144174_at
inx2
Saccharomyces cerevisiae UAS
construct a of Stebbings
2.1
innexin channel activity
Innexin
150746_at
CG12883
2.1
143957_at
CG11971
2.1
nucleic acid binding
Zn-finger C2H2 type
143982_at
Chd3
2.1
ATP dependent DNA helicase ; EC:3.6.1.3
2.1
defense/immunity protein
Mitochondrial glycoprotein
2.1
structural constituent of cytoskeleton
Actin/actin-like, phagocytosis
DEAH-box helicase
154495_at
CG6459
143058_f_at
Act5C
153079_at
CG6550
2.1
144369_at
CG11378
2.1
146611_at
CG6448
2.1
receptor
152631_at
th
thread
2.1
ubiquitin-protein ligase activity, apoptosis
inhibitor activity
Zn-finger RING, Baculovirus inhibitor of apoptosis protein
repeat (BIR)
150834_at
Obp99c
Odorant-binding protein 99c
2.1
PBP_GOBP;odorant binding
Pheromone/general odorant binding protein PBP/GOBP,
Insect pheromone/odorant binding protein PhBP
154669_at
CG5080
146898_at
CG12926
Actin 5C
Deoxyribonuclease/rho motif-related TRAM, Radical SAM,
MiaB-like tRNA modifying enzyme archaeal-type
Protein of unknown function DUF1397
2.1
motor
2.1
tocopherol binding
Cellular retinaldehyde-binding)/triple function, alphatocopherol transport
2.0
transcription factor activity, RNA polymerase II
transcription factor activity
Paired amphipathic helix, H+-transporting two-sector
ATPase
Ferritin 2 light chain homologue
2.0
ferrous iron binding, ferritin;binding
Ferritin/ribonucleotide reductase-like
spastin
2.0
ATPase activity
AAA ATPase central region, MIT, Viral coat and capsid
protein
alpha-tocopherol transfer proteinlike
141716_at
Sin3A
143784_at
Fer2LCH
154205_at
spas
152140_at
CG8678
2.0
enzyme
G-protein beta WD-40 repeat
146771_at
CG1882
2.0
enzyme
Alpha/beta hydrolase fold, Esterase/lipase/thioesterase
149251_at
l(3)04053
2.0
147486_i_at
CG18606
2.0
142778_at
bcn92
142212_at
l(2)08717
2.0
sodium/phosphate cotransporter
Complex 1 LYR protein
2.0
high affinity inorganic phosphate:sodium
General substrate transporter, Major facilitator superfamily
symporter activity, sugar_tr;transporter activity
fer2;electron transporter activity
[2Fe-2S]-binding, Ferredoxin, 2Fe-2S ferredoxin iron-sulfur
binding site
154786_at
CG18522
2.0
147364_at
CG14478
2.0
C-5 cytosine-specific DNA methylase
150582_at
CG13659
2.0
Protein of unknown function DUF227
148144_at
CG14990
2.0
154757_at
CG14991
154136_at
CG31120
mitogen inducible protein-like
serine-type endopeptidase activity,
trypsin;trypsin activity
Peptidase trypsin-like serine and cysteine proteases
2.0
Pleckstrin-like /// IPR009065 // FERM
2.0
2OG-Fe(II) oxygenase superfamily, Prolyl 4-hydroxylase
alpha subunit
146493_at
CG9987
141641_at
cher
cheerio
2.0
2.0
actin binding
143020_at
Talin
talin
2.0
actin binding, cytoskeletal anchoring activity
155055_at
CG31549
2.0
oxidoreductase activity
Short-chain dehydrogenase/reductase SDR, Glucose/ribitol
dehydrogenase
2.0
DNA ligase (ATP) activity
ATP-dependent DNA ligase, BRCT
PDZ/DHR/GLGF domain, Phosphotyrosine interaction
domain, Aminoacyl-tRNA synthetase class I
DNA ligase
Protein of unknown function UPF0027
145021_at
ligase4
151208_at
CG15068
2.0
154462_at
X11L
2.0
enzyme
G-protein coupled receptor activity,
neuropeptide receptor activity
149874_at
CG9918
2.0
141652_at
CG5290
2.0
142203_at
CG14788
152436_at
CG12290
2.0
Rhodopsin-like GPCR superfamily
TPR repeat , TPR-like
2.0
G-protein coupled receptor-like
Filamin/ABP280 repeat, Calponin-like actin-binding, Actinbinding actinin-type
GTP-binding protein HSR1-related, GTP-binding domain
G-protein coupled receptor activity unknown
ligand, rhodopsin-like receptor activity
Rhodopsin-like GPCR superfamily
SINV specific (29 genes)
Probe number
Gene name
142031_at
RpS6
Synomymus
151466_at
CG14599
3.8
150138_at
CG31232
2.8
cyclin-dependent protein kinase regulator
activity
144428_at
CG3603
2.6
oxidoreductase activity acting on CH-OH group Short-chain dehydrogenase/reductase SDR, Rhodopsin-like
of donors, acetoacetyl-CoA reductase activity
GPCR superfamily
ribosomal protein S6
SDV *
8.3
150711_at
CG17189
2.6
149283_at
CG12585
2.5
149489_at
CG7459
151137_r_at
IM4
147027_at
Putative function
copper transporter-like
2.4
copper ion transporter activity
2.3
defense/immunity protein activity
CG30035
2.3
sugar transporter activity
141289_at
CG9821
2.3
142089_at
BG:DS00941.14
150335_at
fit
154079_at
BCL7-like
Prx2540-2
153378_at
Cg25C
149564_at
CG9386
152347_at
CG11841
150052_at
CG31267
144239_at
la costa
148228_at
CG10592
151358_at
CG14189
150743_at
CG31058
147724_at
PIP5K59B
147206_at
L
153731_at
Hsp26
145020_at
CG12175
148888_at
CG4893
146673_at
CG11211
151945_at
RPE65
Cyclin
Odorant binding protein
Immune induced protein 4
146946_s_at
Key words
structural protein of ribosome; small-subunit
cytosol ribosomal protein
Ctr copper transporter
General substrate transporter, Sugar transporter
superfamily
2.3
female-specific independent of
transformer
2.3
Peroxiredoxin 2540
2.2
peroxidase activity, non-selenium glutathione
peroxidase activity
Alkyl hydroperoxide reductase/ Thiol specific antioxidant/
Mal allergen
Collagen type IV
2.2
structural protein
Collagen helix repeat, Collagen triple helix repeat
2.2
BCL7 N-terminal
2.2
serine protease
2.2
serine-type endopeptidase activity
Peptidase trypsin-like serine and cysteine proteases
2.2
serine-type endopeptidase activity
Peptidase trypsin-like serine and cysteine proteases
alkaline phosphatase activity
Alkaline phosphatase
2.2
alkaline phosphatase-like
2.1
2.1
2.1
ligand binding or carrier
1-phosphatidylinositol-4phosphate kinase
2.1
1-phosphatidylinositol-4-phosphate 5-kinase
activity
Phosphatidylinositol-4-phosphate 5-kinase
Lobe
2.1
Heat shock protein 26
2.0
heat shock protein activity
Heat shock protein Hsp20, chaperone
2.0
transcription factor
2.0
C-type lectin-like
P25-alpha
2.0
mannose binding, lectin_c;sugar binding
C-type lectin
2.0
beta-carotene 1515'-monooxygenase activity
Retinal pigment epithelial membrane protein
DCV & FHV specific (39 genes)
Probe number
Gene name
DCV *
FHV *
141374_at
AttA
Attacin-A
8.7
5.2
Gram-negative antibacterial peptide activity
143334_at
scb
scab
7.9
3.5
calcium-dependent cell adhesion molecule activity Integrins alpha chain
146991_at
CG9080
Listericin
7.0
3.6
147421_at
CG14499
5.7
6.7
Diptericin B
5.0
3.7
4.9
2.0
4.7
4.3
147473_at
DptB
147685_at
CG4269
147220_s_at
AttB
147406_at
CG10910
151822_at
Rel
147127_s_at
CG13323
Synomymus
Attacin-B
Relish
Attacin C-terminal region
4.9
antimicrobial peptide activity
Attacin N and C-terminal region
3.1
structural protein
3.5
4.1
specific RNA polymerase II transcription factor
activity
NF-kappa-B/Rel/dorsal
3.2
2.3
Basic helix-loop-helix dimerization domain bHLH
dm
3.2
2.8
DNA binding, transcription factor activity
152356_at
CG32412
3.1
3.5
glutaminyl-peptide cyclotransferase activity
152901_at
CG10383
3.0
2.6
141233_at
CG5966
CG18273
143186_at
h
146364_at
CG5953
141483_at
CG17836
149715_at
mthl5
153722_at
Xpd
141271_at
CG1021
147150_at
Attacin-C
153855_at
mae
147126_i_at
CG32479
148424_at
CG13311
151918_s_at
drpr
154372_at
tamo
141948_at
triacylglycerol lipase
hairy
CG31272
153631_at
CG8675
142932_at
dome
151326_at
CG32666
152225_at
kraken
150549_at
CG11819
141265_at
SH3PX1
153278_at
Cct5
150583_at
153688_at
146100_at
2.8
3.9
2.8
2.4
2.7
3.0
2.7
2.1
Peptidase M28 glutaminyl cyclase
ARM repeat fold
triacylglycerol lipase activity, lipase;catalytic
activity
Esterase/lipase/thioesterase
specific RNA polymerase II transcription factor
activity, specific transcriptional repressor activity
Basic helix-loop-helix dimerization domain bHLH
2.7
2.6
protein dimerization activity, AT_hook;DNA binding HMG-I and HMG-Y DNA-binding domain (A+T-hook)
G protein linked receptorlike
2.7
2.1
G-protein coupled receptor activity
G-protein coupled receptor family 2 (secretin-like)
Xeroderma pigmentosum D
2.7
2.0
helicase activity, general RNA polymerase II
transcription factor activity
ATP-dependent helicase DEAH-box, Helicase c2
2.7
2.7
motor
attacin-like
2.7
4.9
defense/immunity protein
modulator of the activity of
Ets
2.7
3.5
protein binding
2.6
2.5
Ubiquitin-specific protease activity
2.4
2.1
structural molecule activity
Alkaline phosphatase
draper
Replication Protein A 70 replication protein A 70
149642_at
Key words
Attacin N and C-terminal region
antibacterial peptide activity
143126_at
153553_at
diminutive
Putative function
sugar transporter-like
Peptidase C19 ubiquitin carboxyl-terminal hydrolase family
2
2.3
2.1
cell adhesion molecule activity
EGF-like domain, Laminin-type EGF-like domain,
Immunoglobulin/MHC
2.3
2.4
nuclear localization sequence binding
Zn-finger Ran-binding, Immunoglobulin/MHC
2.3
2.0
single-stranded DNA binding
transporter activity
General substrate transporter
protein-tyrosine-phosphatase activity, cytokine
binding
Fibronectin type III, H+-transporting two-sector ATPase
alpha/beta subunit
2.2
2.1
2.2
2.9
domeless
2.1
2.3
2.1
2.5
kraken
2.1
3.1
serine hydrolase activity
Alpha/beta hydrolase, Esterase/lipase/thioesterase
2.1
2.6
protein kinase
C2 calcium/lipid-binding domain CaLB
chaperonin ATPase activity
Chaperonin Cpn60/TCP-1, GroEL-like chaperone ATPase
2.0
3.0
2.0
2.6
CG31436
2.0
2.2
Protein of unknown function DUF227
CG16718
2.0
2.3
Protein of unknown function DUF590
CG13117
2.0
3.9
T-complex Chaperonin 5
SH3, Phox-like
DCV & SINV specific (3 genes)
Probe number
Gene name
145171_at
CG14948
149738_r_at
CG14742
151211_at
CG16844
Synomymus
dpr18
Immune induced molecule 3
DCV *
SDV *
2.2
1.7
2.2
1.7
2.0
1.8
Putative function
Key words
Immunoglobulin-like
defense/immunity protein
FHV & SINV specific (127 genes)
Probe number
Gene name
145820_at
TotM
FHV *
SDV *
Turandot M
Synomymus
47.4
42.4
cathepsin L-like (inactive)
17.0
12.4
9.1
4.6
Putative function
Key words
152626_at
CG11459
144790_at
CG2909
152902_at
CG10341
8.4
2.8
141701_at
CG2064
8.3
4.5
adh_short;oxidoreductase activity
Short-chain dehydrogenase/reductase SDR
149723_at
CG12224
8.1
5.8
oxidoreductase activity
Aldo/keto reductase
151551_i_at
CG30090
serine protease
8.0
6.1
trypsin;trypsin activity
Peptidase trypsin-like serine and cysteine proteases
144460_at
CG6428
asparaginase
7.8
3.3
Lysophospholipase activity
Asparaginase/glutaminase, AMP-dependent synthetase
and ligase
147195_at
CG12505
7.3
5.1
145788_at
CG3008
6.6
3.5
protein kinase activity
Ribulose bisphosphate carboxylase small chain
152598_at
GstE1
Glutathione S transferase E1
6.4
2.5
glutathione transferase activity
Glutathione S-transferase
tribbles
protein serine/threonine kinase activity
Protein kinase
Stress-inducible humoral factor Turandot
cathepsin L activity, Peptidase_C1;cysteine-type
peptidase activity
Peptidase C1A papain
NAF1
Retrotransposon gag protein, Zn-finger CCHC type
149123_at
trbl
6.4
3.5
147139_at
CG10799
6.3
4.0
142206_s_at
CG14527
5.4
4.2
Peptidase_M13_N;metallopeptidase activity
Peptidase M neutral zinc metallopeptidases zinc-binding
site
153894_at
CG11897
5.4
2.6
ABC_membrane;ATP-binding cassette (ABC)
transporter activity
ABC transporter, Nitrogenase component 1 alpha and beta
subunits
149222_at
CG7130
ATP-binding cassette
transporter
5.2
2.4
heat shock protein activity
Heat shock protein DnaJ
150398_at
CG4725
5.1
3.2
Peptidase_M13_N;metallopeptidase activity
Peptidase M13 neprilysin
144845_at
CG15203
5.0
3.0
142545_at
RpA-70
5.0
2.5
single-stranded DNA binding
OB-fold nucleic acid binding domain, Replication factor-A
protein 1
148446_at
Tequila
5.0
4.8
serine-type endopeptidase activity, chitin binding
Peptidase S1 chymotrypsin, Low density lipoproteinreceptor class A, Chitin binding Peritrophin-A
143201_at
ImpL3
4.9
2.1
L-lactate dehydrogenase activity,
ldh;oxidoreductase activity
Lactate/malate dehydrogenase
Replication Protein A 70
Ecdysone-inducible gene L3
155131_at
CG9424
4.8
2.3
151210_at
CG16836
4.6
3.8
Lamino-associated polypeptide 2/emerin
151310_at
CG8620
4.6
4.5
149467_at
CG2791
4.5
2.4
enzyme
153695_at
CG6769
4.5
2.5
transcription factor
147525_at
CG16898
4.5
2.3
Zn-finger C2H2 type
Protein of unknown function DUF227
T-complex protein 1 betasubunit
4.4
3.1
chaperone activity
Chaperonin Cpn60/TCP-1, ATP-dependent helicase DEADbox
Glutathione S transferase D9
4.4
2.3
glutathione transferase activity
Glutathione S-transferase
CG15784
4.4
2.4
CG4721
4.3
2.4
Peptidase_M13_N;metallopeptidase activity
Peptidase M13 neprilysin
4.3
3.0
ATP-binding cassette (ABC) transporter activity
ABC transporter, Ribonucleotide reductase R1 subunit
CG12766
4.0
2.2
trans-12-dihydrobenzene-12-diol dehydrogenase
activity
CG15067
4.0
3.3
142977_at
CG7033
149755_at
GstD9
144521_at
142142_at
154740_at
CG31793
142258_at
147433_at
ATP-binding cassette
transporter-like
Winged helix DNA-binding
Aldo/keto reductase
Cytochrome c
142969_at
Cys
Cystatin-like
4.0
4.0
cysteine protease inhibitor activity
Sarcocystatin, Cysteine protease inhibitor
141583_at
Cyp6a20
cytochrome P450 CYP6A20
3.9
2.1
cytochrome P450 activity
Cytochrome P450, E-class P450 group I
145956_at
CG5958
retinoid binding protein-like
3.9
2.2
retinal binding
150031_at
CG17560
3.9
3.5
142737_at
CG5059
3.9
2.4
147912_at
CG13877
3.7
3.4
153150_at
CG11899
3.7
2.4
phosphoserine transaminase activity,
aminotran_5;transaminase activity
Phosphoserine aminotransferase
141661_at
Ssrp
3.6
2.1
DNA secondary structure binding, single-stranded
RNA binding
HMG1/2 (high mobility group) box
141242_at
CG15092
3.6
2.6
Structure specific
recognition protein
Cellular retinaldehyde-binding)/triple function
Male sterility protein
148051_at
msn
misshapen
3.6
2.1
protein serine/threonine kinase activity, JUN kinase
Serine/threonine protein kinase
kinase kinase kinase activity
153629_at
mre11
meiotic recombination 11
3.6
2.1
endonuclease activity, exonuclease activity
143198_at
Hsp83
Heat shock protein 83
3.5
2.1
chaperone activity, HATPase_c;ATP binding
Heat shock protein Hsp90
155148_at
CG10527
3.5
3.5
farnesoic acid O-methyltransferase activity
Protein of unknown function DM9
148108_at
CG12012
3.5
2.6
144724_at
CG7267
3.5
2.6
151752_s_at
CG33123
leucine--tRNA ligase
3.4
2.1
leucine-tRNA ligase activity, tRNA-synt_1;tRNA
ligase activity
Aminoacyl-tRNA synthetase class Ia, Haem peroxidase
plant/fungal/bacterial
145896_at
CG9508
neprilysin-like
metalloendopeptidase
3.4
2.0
neprilysin activity
Peptidase M neutral zinc metallopeptidases zinc-binding
site
149340_at
rev7
3.4
2.2
cell cycle regulator
DNA-binding HORMA
153982_at
Cct
3.4
2.1
chaperonin ATPase activity
Chaperonin Cpn60/TCP-1, GroEL-like chaperone ATPase
145620_at
CG4259
3.4
2.3
serine-type endopeptidase activity
Peptidase trypsin-like serine and cysteine proteases
152325_at
Lsd-1
3.4
3.4
chaperone
Perilipin
chaperonine-containing Tcomplex gamma-subunit
Lipid storage droplet-1
Metallo-phosphoesterase, DNA repair exonuclease
147353_at
CG11400
3.4
3.1
154266_at
Ada2S
Transcriptional adapter 2S
3.3
2.1
histone acetyltransferase activity
Myb DNA-binding domain, Homeodomain-like
146165_at
CG6113
triacylglycerol lipase-like
3.3
2.9
triacylglycerol lipase activity, sterol esterase
activity
Alpha/beta hydrolase, Esterase/lipase/thioesterase
152063_at
CG4716
3.2
2.5
148973_at
Adgf-A
3.2
2.7
adenosine deaminase activity
Adenosine/AMP deaminase
142599_at
CG31151
3.2
2.2
BAH;DNA binding
Bromo adjacent region
149964_at
CG14872
3.2
2.6
154938_at
CG1583
146000_at
CG7219
150482_at
CG13607
3.1
2.3
142893_at
Rtnl1
3.0
2.7
Reticulon, Calcium-binding EF-hand
150713_at
CG6066
3.0
2.5
Protein of unknown function DUF926
147319_at
CG8317
3.0
2.1
143845_at
Rab6
147564_at
CG16799
145648_at
Cyp309a2
2.9
2.9
cytochrome P450 activity
Cytochrome P450, E-class P450 group I
141815_at
PHGPx
2.9
2.0
glutathione peroxidase activity
Glutathione peroxidase
141688_at
ImpL2
2.9
2.1
cell adhesion molecule activity
Immunoglobulin subtype
Adenosine deaminaserelated growth factor A
serpin
Lipocalin-related protein and Bos/Can/Equ allergen
3.1
2.8
phospholipase A2 activity
Phospholipase A2
3.1
2.5
serine-type endopeptidase inhibitor activity
Serpin
Rab-protein 6
2.9
2.4
RAB small monomeric GTPase activity
lysozyme
2.9
2.4
defense/immunity protein activity, lysozyme activity Glycoside hydrolase family 22
Ecdysone-inducible gene L2
Ras GTPase superfamily
153075_at
CG1773
serine protease-like
2.9
2.3
transmembrane receptor protein serine/threonine
kinase receptor-associated protein activity
Peptidase trypsin-like serine and cysteine proteases
145608_at
CG15358
C-type lectin-like
2.9
2.0
lectin_c;sugar binding
C-type lectin
152320_at
CG6495
2.9
2.1
receptor
Low density lipoprotein-receptor class A
147337_at
CG6429
2.8
2.4
151967_at
CG6415
aminomethyltransferase
2.8
2.1
aminomethyltransferase activity
Glycine cleavage T protein (aminomethyl transferase)
151538_s_at
RfaBp
Retinoid- and fatty-acid
binding protein
2.8
3.0
structural molecule activity, Vitellogenin_N;lipid
transporter activity
von Willebrand factor type D, Lipid transport protein
143197_at
Hsp68
Heat shock protein 68
2.8
2.6
heat shock protein activity
Heat shock protein Hsp70
146311_at
CG9928
2.7
2.1
148547_at
Mocs1
2.7
2.0
152385_at
Aats-gln
150616_at
CG31380
146558_at
151575_at
Glutaminyl-tRNA synthetase
2.7
2.3
2.7
2.3
CG14400
2.7
2.4
CR32218
2.6
2.0
149884_at
CG9616
2.6
3.1
150628_at
CG8932
sodium-dependent
multivitamin transporter-like
2.6
2.0
sodium dependent multivitamin transporter activity Na+/solute symporter
egghead
2.6
2.6
beta-14-mannosyltransferase activity
Peptidylprolyl isomerase FKBP-type
calcium-independent
phospholipase A2
2.6
2.1
calcium-independent cytosolic phospholipase A2
activity
Patatin, Ankyrin
2.5
2.1
2.5
2.1
glycogenin glucosyltransferase activity
2.5
2.0
sepiapterin reductase activity
2.5
2.3
adenosine kinase activity
2.5
2.1
serine-type endopeptidase activity
Peptidase S1 chymotrypsin
2.5
2.3
DNA binding, tRNA_anti;nucleic acid binding
OB-fold nucleic acid binding, Winged helix DNA-binding
hydrogen-exporting ATPase activity
phosphorylative mechanism
Mitochondrial ATP synthase gamma
cathepsin B activity, Peptidase_C1;cysteine-type
peptidase activity
Peptidase C1A papain, Somatomedin B
151468_s_at
egh
142906_at
CG6718
148369_at
CG8209
147607_at
Glycogenin
144701_at
CG12116
152613_at
CG11255
151841_at
CG8586
153194_at
CG9273
146526_at
CG10680
2.5
2.7
144590_at
CG32918
2.5
2.2
146216_at
l(2)06225
2.5
2.1
142954_at
CG5773
2.5
2.3
151839_at
CG31764
2.4
2.1
153218_at
CG3074
2.4
2.3
154405_at
CG11642
2.4
2.2
153673_at
AnnIX
Glycogenin
adenosine kinase
replication factor A 30 kD
subunit
ATP synthase g-subunit
vir-1
Annexin IX
2.4
2.1
glutamine-tRNA ligase activity, ATP binding
Glutamyl-tRNA synthetase class Ic
Protein of unknown function DUF227
motor
Ubiquitin-associated domain, UBA-like, Zn-finger C2H2
type
Glycosyl transferase family 8
Carbohydrate kinase PfkB, Adenosine kinase
Longevity-assurance protein (LAG1), TRAM LAG1 and
CLN8 homology
actin binding, calcium-dependent phospholipid
binding
Annexin type V
142414_at
CG11315
2.4
2.3
152591_at
CG11395
2.4
2.0
146726_at
nec
146136_at
Hand
142768_at
CG7407
153313_at
Rac2
151980_at
149053_at
necrotic
E1 protein and Def2/Der2 allergen
structural protein
2.4
2.3
serine-type endopeptidase inhibitor activity
Serpin
2.3
3.1
transcription factor activity
Basic helix-loop-helix dimerization domain bHLH
2.3
2.2
glass multimer reporter
construct of Nolan
2.3
2.8
small monomeric GTPase activity
Ras GTPase superfamily
CG5150
alkaline phosphatase-like
2.3
2.4
alkaline phosphatase activity
Alkaline phosphatase
CG9451
acid phosphatase
2.2
2.5
acid phosphatase activity
Histidine acid phosphatase
145223_at
CG9059
dipeptidyl-peptidase IV-like
2.2
2.2
dipeptidyl-peptidase IV activity
Peptidase S9 prolyl oligopeptidase,
Esterase/lipase/thioesterase
153434_at
Hn
Henna
2.2
2.2
phenylalanine 4-monooxygenase activity,
ACT;amino acid binding
Aromatic amino acid hydroxylase, Phenylalanine-4hydroxylase tetrameric form
155042_at
scrib
scribbled
2.2
2.2
cell proliferation
Leucine-rich repeat, PDZ/DHR/GLGF domain
147665_at
CG3290
alkaline phosphatase
2.2
2.1
alkaline phosphatase activity
Alkaline phosphatase
144147_at
BM-40-SPARC
SPARC/ostenectin-like
2.1
2.0
ligand binding or carrier
Calcium-binding EF-hand, Osteonectin-like
151851_at
CG6206
alpha-mannosidase
2.1
2.2
alpha-mannosidase activity
Glycoside hydrolase family 38
153583_at
Hsp23
Heat shock protein 23
2.1
2.3
heat shock protein activity
Heat shock protein Hsp20
145890_at
CG9498
2.1
2.0
150030_at
CG17562
2.1
2.2
enzyme
147336_at
CG6426
calcium binding protein-like
2.0
2.9
ligand binding or carrier
Destabilase, Calcium-binding EF-hand
144903_at
CG10352
4-nitrophenylphosphataselike
2.0
2.0
4-nitrophenylphosphatase activity
Haloacid dehalogenase-like hydrolase
153365_at
CG15825
2.0
2.1
143917_at
lush
lush
2.0
2.4
odorant binding
Insect pheromone/odorant binding protein PhBP
145598_at
CG17646
152488_at
CG8343
ATP-binding cassette
transporter
* Fold induction compared to Tris injection control
Protein of unknown function DUF227
Male sterility protein
2.0
2.5
ATP-binding cassette (ABC) transporter activity
ABC transporte, AAA ATPase
2.0
2.0
mannose binding, lectin_c;sugar binding
C-type lectin
Supplemental Table III: List of the 42 genes up-regulated by DCV, FHV and SINV
Probe number
153941_at
153231_at
146285_at
145307_at
146814_at
144846_at
141626_at
141353_at
Gene name
Fst
CG8791
CG31764
CG15043
CG12780
CG2081
CG7635
CG6687
150837_at
Obp99b
152122_at
144998_at
152721_at
153233_at
144882_at
147430_at
146256_at
149782_at
149477_at
152112_at
152183_at
CG10912
CG15745
Idgf1
CG6357
PGRP-SA
CG15066
CG14934
Hsp70Bc
CG11671
CG13625
CG4680
142415_at
Nmda1
143682_at
149139_at
141511_at
Tig
CG4786
dos
151767_at
Nmdmc
154760_at
CG4618
146873_at
154821_at
153197_at
153527_at
143005_at
150036_at
154140_at
146708_at
144068_at
143802_at
147410_at
141685_at
143401_at
152618_at
148843_at
155151_at
Cyp4p3
CG8147
Eip75B
CG1572
CG4199
CG14907
CG4713
Tsp42Ed
IM2
CG14906
CG10911
CG3066
Uro
Tsp42El
CG13075
CG10916
Synomymus
Frost
Fold induction
SINV E. c
M. l
2.9
16.9 23.8
-4.8
2.5
3.5
4.6
4.1
3.1
2.1
3.6
4.7
1.6
1.9
3.1
1.0
0.9
4.7
2.2
2.7
2.9
6.5
2.7
5.0
B. b
8.6
1.0
1.9
3.9
2.2
1.0
4.0
7.4
L. m
15.5
4.5
3.7
2.3
2.7
1.5
2.5
5.7
GNBP-like
Vago
Mec2
Serpin 88Eb
DCV
19.7
7.5
7.1
6.9
6.5
5.3
4.8
4.5
FHV
10.9
3.7
9.5
7.3
5.4
6.0
2.9
10.3
Odorant-binding protein 99b
3.8
10.8
4.6
0.2
0.2
0.2
0.3
3.6
3.4
3.4
3.3
3.2
3.1
3.1
3.1
3.1
2.9
2.9
7.7
10.9
3.2
4.9
3.9
2.2
7.7
5.5
5.0
2.4
13.7
2.3
8.5
2.8
2.6
2.9
3.2
3.7
5.4
2.0
2.1
8.6
1.6
2.3
2.3
1.3
3.6
7.8
1.0
0.7
1.1
1.3
0.8
1.8
2.7
2.0
1.7
3.6
16.4
1.6
8.2
1.5
1.5
0.9
2.4
2.7
1.8
2.0
2.6
19.0
1.0
2.0
2.3
2.1
1.4
2.7
2.5
2.1
2.5
7.5
1.6
1.1
4.8
2.1
1.0
3.1
Putative function
Respond to cold. Stress response.
High affinity inorganic phosphate:sodium symporter activity
Unknown, response to virus
Unknown
Gram-negative bacterial cell surface binding
Unknown, antiviral activity
Unknown, Band 7 protein; Stomatin
Serine protease inhibitor
Autophagic cell death. Pheromone/general odorant binding
protein
Protein of unknown function DUF725
Unknown
Chitinase II; Glycoside hydrolase, family 18
Cysteine-type endopeptidase activity.
Peptidoglycan recognition
Unknown
Alpha amylase; Glycoside hydrolase,
Heat shock protein 70
Unknown
Unknown
Unknown
Vir-1
Imaginal disc growth factor 1
IM23
Mal-B1 (Maltase B1)
N-methyl-D-aspartate receptor-associated
protein
Tiggrin
Rcd2 (Reduction in Cnn dots 2)
daughter of sevenless
NAD-dependent methylenetetrahydrofolate
dehydrogenase
CHMP2B (Charged multivesicular body
protein 2b)
Ecdysone-induced protein 75B
l(2)gd1 (lethal (2) giant discs 1)
Tetraspanin 42Ed
Immune induced molecule 2
Sp7 (Serine protease 7)
Urate oxidase
Tetraspanin 42El
2.7
4.9
2.9
1.8
1.7
1.6
1.7
N-methyl-D-aspartate selective glutamate receptor activity
2.6
2.6
2.5
3.9
3.2
4.9
3.4
2.5
2.8
1.2
8.8
2.2
1.8
8.1
3.2
2.0
2.7
2.9
1.7
1.5
1.0
2.5
5.4
5.1
2.8
3.6
0.8
1.3
Haem oxygenase-like, multi-helical.
Unknown, von Willebrand factor, type C
Regulation of Ras protein signal transduction
NAD(P)-binding domain; Tetrahydrofolate dehydrogenase/
cyclohydrolase
2.5
2.5
2.3
1.7
2.7
2.9
1.8
Unknown
2.5
2.5
2.5
2.4
2.4
2.4
2.4
2.4
2.4
2.3
2.3
2.2
2.2
2.1
2.1
2.0
2.1
5.1
3.3
2.1
5.8
5.4
3.3
4.0
2.6
4.4
2.9
4.1
2.2
4.0
3.1
6.7
2.0
4.6
2.1
2.0
4.5
2.6
2.3
2.5
2.7
2.8
2.3
3.0
2.1
2.2
2.2
4.3
3.4
1.0
1.3
1.1
2.3
2.3
0.8
1.5
2.7
1.3
1.4
1.9
3.8
0.8
4.5
1.3
4.8
0.3
1.3
1.2
2.5
2.1
1.2
1.4
4.1
1.3
1.4
2.0
5.0
0.9
5.8
1.2
3.6
1.0
1.8
1.1
1.8
2.7
1.6
2.0
3.5
1.7
1.5
1.8
5.5
1.4
3.8
1.7
2.0
0.4
2.7
0.8
1.4
2.7
1.2
2.2
1.4
0.9
1.2
4.1
2.0
1.5
4.7
2.1
Cytochrome P450, E-class, group I
Alkaline phosphatase-like
Nuclear hormone receptor,
Unknown, MARVEL-like domain
FAD/NAD-linked reductase
Unknown
C2 calcium-dependent membrane targeting
Unknown
Unknown
Methyltransferase activity
Protein of unknown function DUF725
Clip domain chymotrypsin-type peptidase
Urate oxidase activity
Unknown
Chitin binding
Zinc ion binding
Upregulation superior to 2-fold is shown in bold. Data for Escherichia coli (E.c), Micrococcus luteus (M.l) and
Beauveria bassiana (B.b) are from Ref 19. Data for Listeria monocytogenes (L.m) are from Ref 30.