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PERSPECTIVE
GATA-Binding Transcript ion Factors in Hema to poie t ic Cells
By Stuart H. Orkin
D
IFFERENTIATION of hematopoietic cells involves
progressive restriction of developmental potential as
pluripotent hematopoietic stem cells give rise to multipotential progenitors that subsequently mature along single
lineages. Although mechanisms responsible for decisionmaking in hematopoietic development are not defined, it is
likely that they involve two distinct, but interrelated, pathways: one involving nuclear regulatdry proteins that directly
control and coordinate gene expression, and a second
involving signal transduction from growth factors through
their cognate surface receptors. One goal of research
efforts is identification of lineage-restricted nuclear regulatory proteins that establish the unique and characteristic
gene expression profiles of mature hematopoietic cell types.
In principle, searches for such factors might rely on finding
cell-specific DNA-binding proteins characterized by the
presence of sequence motifs found in known regulators in
other systems (such as homedomains, zinc fingers, helixloop-helix, or basic zipper motifs) or on discovering novel,
putative transcription factor genes at the sites of chromosomal translocations in leukemias. An inherent difficulty
with these approaches relates to ignorance regarding target
genes acted on by these proteins and, hence, in appreciating
the roles of putative regulators during normal cellular
differentiation. The identification of nuclear DNA-binding
proteins that recognize short DNA sequences in cisregulatory elements associated with lineage-specific transcription offers an alternative and more direct route to
relating controlling proteins and target genes. If proteins
identified in this manner regulate sets of genes characteristic of a lineage, an understanding of the mechanisms of
transcriptional control of the regulator itself provides a
means of unravelling the first steps in the pathway of
lineage development.
From this perspective, I will review evidence implicating
transcription factors of a small family, the GATA-binding
proteins, in control of gene expression and development of
hematopoietic cells. Questions raised by recent studies are
considered in relation to approaches that may provide
definitive answers in the future.
genes with GATA-motifs in defined cis-elements is likely to
extend well beyond hematopoietic cells. A tabulation of
genes with GATA-motifs of functional relevance is presented in Table 1. To avoid inclusion of genes containing
irrelevant GATA sequences, the examples listed in the
table are limited to those where the role of the element has
been tested experimentally in a transfection assay or where
other data support the relevance of the site. Thus, this
listing is representative, rather than comprehensive.
GATA, as a cis-element, was first defined in analyses of
globin genes. Evans et ai2 reported that ail chicken globin
gene promoters contained a GATA-sequence conforming
to the consensus [T/A(GATA)A/G]. Wall et a13 noted the
presence of multiple GATA-motifs in the minimal human
3’-p-globin gene enhancer. Martin et ai4 demonstrated that
a GATA-element in the human y-globin gene promoter is
necessary for overexpression consequent to a mutation
causing the hereditary persistence of fetal hemoglobin
syndrome. Studies by other groups amplified these initial
studies implicating GATA motifs in globin gene expression.5-10
The finding that GATA-elements are required for full
promoter activity of other erythroid-expressed, nonglobin
genes, such as porphobilinogen deaminase (PBG-D),11,12
immediately suggested a broader role in erythroid cells.
Indeed, at present there are no erythroid-expressed genes
that have been shown to be independent of GATA-motifs
within their promoters or enhancers. This generalization
applies to membrane proteins, the erythroid lineage growth
factor receptor (erythropoietin receptor, EpR),13-15 and
transcription factors, such as GATA-116 and SCL (tal-l)17
(see below).
A potentially significant aspect of the role of GATAelements relates to the variety of locations in which they
may be found and to variations in GATA-motif sequences.
In addition to their presence within promoters and enhancers of erythroid-expressed genes, GATA-motifs are found
in the active core regions of the locus control regions
( L C R S ) ~of~the
. ~ ~human OL- and P-globin gene cluster^.^-^^
THE GATA-MOTIF AS A cis-REGULATORY ELEMENT
From the Division of Hematology /Oncology, Children’s Hospital
and the Dana Farber Cancer Institute, the Department of Pediatrics,
Harvard Medical School, Howard Hughes Medical Institute, Boston,
MA.
SubmittedApril24, 1992; accepted April 24, 1992.
Supported in part by grants from the National Institutes of Health.
S.H.O. is an Investigator of the Howard Hughes Medical Institute.
Address reprint requests to Stuart H. Orkin, MD, Depament of
Hematology, Children’s Hospital, 300 Longwood Ave, Boston, MA
02115.
The publication costs of this article were defrayed in part by page
charge payment. This article must therefore be hereby marked
“advertisement” in accordance with 18 U.S.C. section 1734 solely to
indicate this fact.
0 1992 by TheAmerican Society of Hematology.
0006-4971/92/8003-0032$3.00/0
Short DNA sequences within control elements of genes
constitute cis-regulatory motifs recognized by nuclear proteins that participate in transcription.’ These include (among
many others) motifs such as TATA, which provides the site
for binding of TFIID as part of the basal transcriptional
machinery, and CCAAT, which is an important element in
many gene promoters. Through studies largely focused on
genes expressed in hematopoietic cells, the motif GATA
has become recognized as a cis-regulatory element in
diverse genes. Although its potential relevance first became
apparent in the context of gene expression in erythroid
cells, genes expressed in other hematopoietic lineages and
endothelial cells also use this motif. In fact, the range of
Blood, VOI 80, NO3 (August l), 1992: pp 575-581
575
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STUART H. ORKlN
576
Table 1. GATA-Binding Sites in cis-Regulatory Elements
1. Erythroid-expressed genes and globin regulatory sequences
Globin genes:
Promoters: either proximal at the TATA-box region (eg, chicken
pA) or within the promoter (numerous examples in chicken,
human, mouse, frog)
Enhancers: 3’-p-globin (human, chicken); 3’-y-globin (human)
Locus control regions (LCRs):
p-LCR: all active core regions
wLCR
Transcription factor genes:
GATA-1 promoter (mouse, human, chicken)
Stem cell leukemia (SCL, tal-1) promoter
Growth factor receptor genes:
Erythropoietin receptor (EpR) promoter (human, mouse)
Other genes:
Heme biosynthetic enzyme gene promoters (eg, porphobilinogen
deaminase, PBG-D), pyruvate kinase, glycophorin
2. Megakaryocytic-expressedgenes:
Platelet factor 4 promoter (rat), platelet glycoproteins Ilb, Ib
promoters (Gpllb, Gplb)
3. Mast cell-expressed genes:
Mast cell carboxypeptidase A (MC-CPA)promoter
4. Endothelial cell-expressed genes:
Preproendothelin-I promoter
Vascular cell adhesion molecule-1 (VCAM-1)
LCRs have the distinctive property of insulating genes from
chromosomal position effects and providing a favorable
context for their expression in vivo.26As such, globin genes
in cis to LCRs are expressed at a high level in an erythroid
environment, independent of integration site. The overall
activity of LCRs on linked gene expression appears to be
dependent on three core DNA motifs: GATA, CACCC
(GGTGG), and TGAGTCA (an AP-1m ~ t i f ) . ~ -Integra*~.~
tion site independence is best attributed to an interaction of
GATA and CACCC motifs (and their bound proteins).
Thus, the GATA-motif is used in a variety of settings:
promoters, either upstream or near to the start site of
transcription initiation; enhancers, usually located 3’ in the
globin genes; and LCRs, situated far upstream in the
human globin gene clusters. Moreover, the precise sequence of GATA-motifs varies considerably, particularly
with respect to the nucleotides flanking the GATA-core. In
addition, GATA-motifs are sometimes present in more
complex arrangements, such as when they are spaced
closely together or are o ~ e r l a p p i n g .Although
~ ~ , ~ ~ ~the
~ full
import of these variant motifs is not certain, it is suspected
that they herald a different functional context in vivo. These
observations suggest that proteins interacting with GATAmotifs may be versatile in their functions. As noted below,
recent data support this c0ncept.2~
As discussed below, studies of nuclear proteins binding
to the GATA-motif led to the recognition that promoters
for megakaryocytic and mast cell-expressed genes also
contain GATA-motifs that are critical for promoter activity
in transfection as~ays.30-3~
In addition, the functional interactions of GATA-elements with enhancers may be complex, as suggested by dissection of the megakaryocytic rat
PF4 promoter.33 Preliminary evidence suggests that the
GATA-motif may be particularly prominent in the promoters for mast cell-specific
Use of the GATA-motif is not restricted to genes
expressed only in hematopoietic cells. Studies of the endothelial-expressedpreproendothelin-1 gene showed a GATAsite that was essential for promoter f~nction.3~
Moreover,
recent work on VCAM-1 implicates a double GATA-site in
promoter function and response to inflammatory mediators
(T. Collins, personal communication).
GATA-BINDING PROTEINS
Transcriptional effects of cis-regulatory elements are
mediated through the binding of sequence-specific nuclear
proteins.’ The cell is provided with a formidable challenge
in achieving transcriptional specificity endowed with proteins that recognize only short target DNA sequences (on
the order of 6 to 8 bp). Transcriptional specificity depends
on multiple factors, including the availability of specific
DNA sequences in native chromatin and the interaction of
DNA-binding proteins with neighboring binding-proteins,
as well as proteins that do not directly contact DNA.
Furthermore, canonical cis-regulatory motifs are often
recognized (at least in vitro) by several proteins, which may
co-exist in nuclear extracts of cells. On occasion these
proteins may be derived from different protein classes (such
as helix-loop-helix and zinc finger proteins).
Four years ago, when GATA as a motif was first
identified in globin gene promoters and enhancers, the
situation appeared deceptively simple: a single, abundant
erythroid cell nuclear pr0tein,3~,~~
now known as GATA-1,19
was observed to bind the sequence with high affinity.
Through cDNA cloning and cross-hybridization screening,
independent but related proteins, referred to as GATA2,3,19,37
and 4 (T. Evans, D. Wilson; unpublished data), have
been characterized. Each exhibits a characteristic pattern
of expression during development (see below) and binds
specifically to the GATA-motif.
Members of the GATA-binding protein family are related by virtue of a highly conserved protein domain that is
necessary and sufficient for DNA-recogniti~n.~’DNAbinding domain is conferred by two homologous, finger
domains whose architecture resembles zinc-fingers of the
cys-cys variety. As the fingers of the GATA-binding proteins are distinctive in their organization, it is likely that
they represent a new family of zinc-finger proteins with a
novel structure. Consistent with this view is the existence of
a very similar finger structure in
Drosophila (D.
Engel, S-F. Tsai; unpublished data), and C e l e g a n GATA~~~
binding proteins. Curiously, a single-finger domain is capable of binding to a GATA-consensus in the fungal proteins,
whereas two fingers are found in Drosophila, C elegans, and
vertebrate members of the family. An interesting question
posed by these findings is “do two fingers have advantages
over one” in DNA recognition or transcriptional activation
of target genes?
Conservation of primary amino acid sequence among the
finger regions of the GATA-binding proteins of mammals,
avians, and amphibians is remarkable.“‘I Selected amino
acid differences are characteristic of the different GATAproteins, such that the fingers of GATA-1 can be distinguished from GATA-2 and 3, and similarly GATA-2 and
GATA-3 can be distinguished from each other and from
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GATA-BINDING TRANSCRIPTION FACTORS
577
GATA-1.40 The biologic significance of such differences is
unknown.
Transcription factors are modular proteins.’ In general,
DNA-binding domain(s) are separable from regions that
mediate interactions with other proteins and thereby activate transcription. This division of labor is also evident in
the GATA-binding proteins. Using simple reporter constructs containing a minimal promoter plus a GATAbinding site, transcriptional activation by GATA-binding
proteins has been demon~trated.”.~~
Among the GATAproteins, GATA-1 and GATA-4 are potent activators,
whereas GATA-2 and GATA-4 are more modest. Activation of transcription in transfected fibroblasts is dependent
on domains of the proteins located outside the finger
regions. In mouse GATA-1 qualitatively different domains
have been identified by fusion of N- and C-terminal regions
to a heterologous DNA-binding d0main.2~The N-terminal
domain confers transcriptional activation in this assay,
whereas the C-terminal portion does not.
Comparison of the primary amino sequences of the
GATA-binding protein family members highlights changes
that may be of functional significance. The evolutionary
relationships of GATA-1, 2, and 3 of mammalian, avian,
and amphibian origins and the fungal protein are represented in Fig 1. Outside their DNA-binding domains,
GATA-2 and GATA-3 are more similar to each other than
to GATA-1. Conservation of GATA-2 and GATA-3 is
strongly maintained among avians, amphibians, and mamareA
XGATA- 1
cGATA- 1
mGATA- 1
hGATA-I
xGATA-3
cGATA-3
mGATA-3
hGATA-3
hGATA-2
xGATA-2
cGATA-2
Evolutionary time
Fig 1. Evolutionary relationships of protein of the GATA-binding
family. Primary amino acid sequences were compared using the
PILEUP program of the Genetics Computer Group (University of
Wisconsin, Biotechnology Center, Madison). Note that xGATA-1 and
c-GATA-1divergefrom mammalianGATA-1 at an earlier point and are
divergentfrom each other. GATA-2 and GATA-3 divergence is of more
recent origin. Species: x Wenopus),c (chicken). m (mouse), h (human).
Table 2. Cellular Distribution of GATA-Binding Proteins
Family
Member
GATA-1
GATA-2
GATA-3
Sites of Expression
Erythroid, mast, megakaryocytic lineages
Multipotential progenitor cells and lines
Mast, megakaryocytic lineages
Early erythroid cells (chicken)
Multipotential progenitor cells and lines
Endothelialcells
Embryonic brain
Various others (including ES, myeloid cells)
T-lymphoid cells
Embryonic brain
Various others (including kidney, endothelial, and
ES cells; chicken red blood cells)
mal^?^,^ In contrast, GATA-1 proteins have undergone
appreciable divergence over evolutionary time.4O Although
the mammalian proteins are highly conserved (save for
conservative amino acid replacements), avian and amphibian primary sequences are divergent from each other and
from mammals (see top portion, Fig 1). This is provocative
for two reasons. First, important transcription factors are
generally highly conserved across species. Second, because
the salient features of globin gene regulation appear similar
in all vertebrates, it is surprising that GATA-1, a factor
essential for erythroid development (see below), differs
greatly among species. The biologic significance of such
divergence is not yet apparent, but should be the focus of
future investigation.
Recent studies suggest that GATA-1 is versatile in its
functions and may serve to mediate an interaction between
the 3’-enhancer and a noncanonical TATA-box in the
chicken P-globin gene.29 Thus, depending on where it
binds, GATA-1 may serve alternatively as a direct activator
of transcription (presumably through interaction with TFIID
or other components of the basal transcriptional machinery) or as a mediator of promoter-enhancer activity. It is
quite likely that interactions between LCR elements and
individual globin genes may be mediated by GATA-1 in a
manner similar to that described for the chicken P-globin
promoter and enhancer.
CELLULAR DISTRIBUTION OF GATA-BINDING PROTEINS
Each GATA-binding protein exhibits a distinctive pattern of expression in tissues and cell lines, which is broadly
summarized in Table 2. Expression of GATA-1, the socalled “erythroid transcription factor,” is hematopoietic
specific and abundant in erythroid, megakaryocytic, and
mast cell l i r ~ e a g e s . ~Expression
~ , ~ ~ , ~ ~in. ~these
~ three related
hematopoietic lineagesimplies initiation of GATA-1expression in a multipotential progenitor cell and maintenance of
its expression thereafter, or independent activation of
GATA-1 expression after formation of each lineage. Evidence favors the first possibility, as GATA-1 RNA (and
protein) is detected in multipotential mouse cell lines, as
well as purified human CD34+ progenitors and their
immediate d e c e n d a n t ~ .GATA-1
~ ~ ~ ~ ~ is not appreciably
expressed in lymphoid cells, monocytelmacrophages, or
neutrophilic cells (differentiated from cultured HL60 cells).
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578
STUART H. ORKlN
Hence, if GATA-1 is expressed in the colony-forming
unit-granulocyte macrophage (CFU-GM) multipotential
progenitor, its expression must be downregulated during
myeloid differentiation. Indeed, data with permanent mouse
cell lines and differentiated, human CD34+ progenitors
support this model.41Therefore, we currently envision that
GATA-1 is first expressed during hematopoietic development at the multipotential CFU-GM stage, and then
subsequently upregulated during erythroid maturation and
downregulated along the myeloid pathway.
The other vertebrate GATA-binding proteins are expressed in different profiles. GATA-2 RNA is abundant in
megakaryocytes, mast cells, and interleukin-3 (IL-3)dependent mouse hematopoietic cell lines, but low or
undetectable in mouse erythroleukemia cell line^.'^,^^
GATA-2 RNA is also expressed in primitive chicken
erythroblast^.^^ Protein levels in cells or tissues are unknown, as specific antisera have not yet been developed.
Outside the hematopoietic system, GATA-2 is expressed at
low levels in many cell types and at higher levels in selected
tissues. Prominent expression is seen in the embryonic
nervous system in vertebrates (D. Engel, L. Zon; unpublished data), perhaps suggestive of a critical role for this
factor in neuronal development.
GATA-3 is highly expressed in T-lymphoid cells of all
Taken together with the identification of
functionally important GATA-motifs in the T-cell receptor
a-and 6-chain enhancers, it is likely that GATA-3 serves as
a transcriptional activator for T-cell-specific genes. GATA-3
is also expressed at high level in developing brain, although
in a slightly different distribution from that of GATA-2 (D.
Engel, L. Zon; unpublished data). Hence, GATA-2 and
GATA-3 may have distinctive roles in development of the
nervous system. Finally, GATA-3 is expressed widely at low
levels in cell lines (unpublished data).
GATA-4, the newest member of the family, is expressed
in a very limited tissue distribution in the developing mouse
and frog (T. Evans, D. Wilson; unpublished data). Precise
identification of the expressing cell types is under study.
ROLES OF GATA-BINDING PROTEINS IN
HEMATOPOIETIC CELLS
The mere presence of a transcription factor within a cell
does not ensure its role in gene expression or development.
The extent to which important cellular functions and
pathways are protected by functional redundancy may be
extensive.46A powerful approach to assigning the roles of
specific proteins in developmental pathways uses homologous recombination to generate targeted mutations of
chosen genes in pluripotent, mouse embryonic stem (ES)
~ e l l s .After
4 ~ injection into host blastocysts, ES cells contribute to somatic tissues and the germline. Hence, the phenotype referable to specific mutations may be shown. Furthermore, the potential for hematopoietic development from
ES cells can also be evaluated directly in
Through gene targeting in ES cells it has been established that GATA-1 is, indeed, essential for erythroid
development of both primitive and definitive lineages.50As
GATA-1 is located on the X-chromosome,sl disruption of
the gene in male ES cells generates a null with respect to
GATA-1 expression, and obviates the need to pass the
mutation through the mouse germline to uncover its consequence. GATA-1 null ES cells do not contribute to the
mature erythroid cells of chimeric animals.5O Consistent
with this, null ES cells fail to differentiate into erythroid
cells in vitro and do not express any embryonic globin
RNAs.52This phenotype is correctable in vitro by transfer
of the normal GATA-1 into the null ES cells.52
However, hematopoietic cells lacking GATA-1 do give
rise to mature myeloid cells in vivo and in ~ i t r o . ~Hence,
~,~*
the block to differentiation in GATA-1-deficient cells is at
a later stage than the hematopoietic stem cell or its
immediate descendents. In fact, hematopoietic cells destined for erythroid maturation are arrested at the proerythroblast stage.52 Whether this block reflects inadequate
expression of the EpR gene, which is at least partially
regulated through GATA-1,I3-l5or failure to express another critical product is unknown. The potential effects, if
any, of GATA-1 deficiency on megakaryocytic and mast cell
development are under study. These experiments define
GATA-1 as an essential erythroid transcription factor, and
establish a paradigm for the genetic analysis of other genes
involved in hematopoietic development.
What roles GATA-2 and GATA-3 may play in hematopoietic cells remain speculative. Abundant expression of
GATA-2 RNA in megakaryocytes and mast cells raises the
possibility that it, rather than GATA-1, regulates genes
specifically in these lineages. Furthermore, high expression
of GATA-2 in multipotential, IL3-dependent progenitor
cell lines suggests possible involvement in hematopoiesis
before the requirement for GATA-1. Abundant expression
of GATA-3 in T-lymphoid cells and the identification of
important GATA-elements in T-cell receptor enhancers
predict a role for this factor in lymphoid development.
Outside the hematopoietic system the expression of
GATA-2 and GATA-3 in embryonic brain suggests potential roles in neuronal development. Finally, GATA-2 is
likely to serve an important function in transcriptional
control of several endothelial-expressed genes. Many of
these predictions should be considered tentative until the
consequences of targeted mutation of GATA-2 and
GATA-3 on development become known. It may only be
through such genetic approaches that the selective roles of
these proteins will be clarified.
EARLY EVENTS AND COMMITMENT IN HEMATOPOIESIS:
GATA-1 GENE REGULATION AND COUPLING TO OTHER
TRANSCRIPTION FACTORS
An important goal of studies of transcriptional regulators
of cell-specific gene expression is an understanding of the
mechanisms involved in initiating a program (or cascade) of
development. Insights might be derived from two experimental inqueries: first, how are GATA-binding proteins, notably GATA-l, regulated during hematopoietic development? Second, what regulatory factors might be under the
control of GATA-1 (or its relatives)?
As noted above, GATA-1 is thought to be expressed in
multipotential progenitors cells and then either upregu-
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GATA-BINDING TRANSCRIPTION FACTORS
579
lated in erythroid development or downregulated in myeloid mat~ration!~.~~
Analyses of the GATA-1 gene promoter have defined an upstream double GATA-motif that
is required for full activity and is bound in vivo in native
erythroid chromatin.16 Thus, GATA-1 appears to participate in positive regulation of its own promoter. Autoregulation may serve to maintain the differentiated state. While
these data suggest how upregulation of GATA-1 in erythroid development may be achieved, they fail to account
for its initial expression. Elucidation of the factors responsible for the onset of expression will provide novel insights
into the pathway of hematopoietic development.
Existing evidence indicates that GATA-1 plays an important role in regulation of the EpR promoter; however, it
cannot be the sole factor responsible for its expression.13-15
Nonetheless, involvement of GATA-1 in EpR expression is
a mechanism by which viability and subsequent development of erythroid progenitors may be guaranteed. If multipotential progenitors display a variety of lineage-restricted growth factor receptors at low level, chance
encounter with Epo could “select” erythroid progenitors
for further maturation. Positive influences of GATA-1 on
its own expression and on that of the EpR would, in
principle, cooperate to promote erythroid development.13-15
As an early transcription factor in erythroid development, it is of interest to ask whether GATA-1 regulates
other transcription factors, which might be placed downstream in a regulatory hierarchy. One potential target gene
for GATA-1 (or GATA-2 action) is that encoding the
helix-loop-helix protein, SCL (stem cell leukemia) or tal1.53~5~
The SCL gene, first discovered at the site of translocations in T-cell acute lymphoblastic leukemia, is normally
expressed in erythroid, mast, and megakaryocytic lineages.
This profile is remarkably similar to that of GATA-1 itself.
Structurally, SCL can be placed in the basic-HLH class of
transcription factors, of which my0D,5~an important myogenic regulatory protein, is a member. In its proximal
promoter, the SCL gene displays a GATA-motP that
binds GATA-1 (or GATA-2) with high affinity, is required
for full promoter activity in transfection experiments, and
mediates transactivation of reporters by overexpressed
GATA-protein (Aplan PD, Nakahara K, Orkin SH, Kirsch
I R in preparation). Hence, at this rudimentary level, SCL,
a presumed erythroid transcription factor by analogy to
myoD and other basic-HLH proteins, may be one target for
GATA-1 (or GATA-2) action. If this proves to be the case,
it relates two transcription factor families in hematopoiesis
and may begin to establish an order of regulatory events in
hematopoietic development.
UNRESOLVED ISSUES
A firm role in hematopoietic development for one member of the GATA-binding family of transcription factors,
GATA-1, has been established. Although GATA-1 is
essential for erythroid development, little is understood
regarding its interactions with other proteins that cooperate to activate erythroid gene expression, mediate the
function of LCRs, or switch globin genes during ontogeny.
If GATA-1 is (as it would appear) a master regulator of the
end-stage products of erythroid maturation, why is it that
these markers are not expressed in mast cells and megakaryocytes where GATA-1 is also abundant? Perhaps there are
cell-specific protein interactions with GATA-1 that determine the sets of genes activated in each lineage. What is the
role, if any, of GATA-2 in early hematopoietic development, or in mast or megakaryocytic gene expression? Does
GATA-3 fulfill its anticipated role as an important factor
for T-lymphoid cell development? Is there cross-talk between members of the GATA-binding protein family during development, such that one controls expression of
another? What environmental cues provided by growth
factors lead to the selective activation or repression of these
proteins in development? How are these proteins linked to
the elusive issue of commitment of progenitors to individual
lineages during hematopoietic restriction?
Recent progress in the use of genetic approaches to
complement conventional molecular approaches to transcriptional regulation suggests that answers to many of
these questions will be forthcoming in the future. As other
transcription factors that play selective roles in hematopoietic cells are identified, regulatory hierarchies and interactions between classes of regulators may be shown. As the
trajectory of this research converges with signal transduction through growth factor receptors, we may come to
understand in mechanistic terms how hematopoietic lineages are chosen during development.
ACKNOWLEDGMENT
I am grateful to the members of my laboratory (past and present)
and our collaborators, who have contributed to studies of the
GATA-binding protein family, for valuable discussion and criticism.
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Activation of the erythropoietin receptor promoter by transcription factor GATA-1. Proc Natl Acad Sci USA 88:10638,1991
14. Heberlein C, Fischer K-D, Stoffel M, Nowock J, Ford A,
Tessmer U, Stocking C The gene for erythropoietin receptor is
expressed in multipotential hematopoietic and embryonal stem
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15. Chiba T, Ikawa Y, Todokoro K GATA-1 transactivates
erythropoietin receptor gene, and erythropoietin receptor-mediated signals enhance GATA-1 gene expression. Nucleic Acids Res
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16. Tsai S-F, Strauss E, Orkin SH: Functional analysis and in
vivo footprinting implicate the erythroid transcription factor
GATA-1 as a positive regulator of its own promoter. Genes Dev
5:919,1991
17. Aplan PD, Begley CG, Bertness V, Nussmeier M, Ezquerra
A, Coligan J, Kirsch IR: The SCL gene is formed from a
transcriptionallycomplex locus. Mol Cell Biol 10:6426, 1990
18. Grosveld F, van Assendelft GB, Greaves DR, Kollias B:
Position-independent, high-level expression of the human betaglobin gene in transgenic mice. Cell 51:975,1987
19. Orkin SH: Globin gene regulation and switching: Circa 1990.
Cell 63:665,1990
20. Talbot D, Grosveld F The 5’ HS 2 of the globin locus
control region enhances transcription through the interaction of a
multimeric complex binding at two functionally distinct NF-E2
binding sites. EMBO J 10:1391,1991
21. Talbot D, Philipsen S, Fraser P, Grosveld F Detailed
analysis of the site 3 region of the human P-globin dominant
control region. EMBO J 9:2169,1990
22. Strauss EC, Orkin SH: Human P-globin locus control region
hypersensitive site 3: In vivo protein-DNA interactions. Proc Natl
Acad Sci USA 1992 (in press)
23. Strauss EC, Andrews NC, Higgs DR, Orkin S H In vivo
footprinting of the human a-globin locus upstream regulatory
element by guanine/adenine ligation-mediated PCR. Mol Cell Biol
12:2135,1992
24. Jarman AP,Wood WG, Sharpe JA, Gourdon G, Ayyub H,
Higgs DR: Characterization of the major regulatory element
upstream of the human a-globin gene cluster. Mol Cell Biol
11:4679,1991
25. Philipsen S, Talbot D, Fraser P, Grosveld F The @-globin
STUART H. ORKlN
dominant control region: Hypersensitive site 2. EMBO J 9:2159,
1990
26. Felsenfeld G: Chromatin as an essentialpart of the transcriptional mechanism. Nature 355:219,1992
27. Martin D, Orkin S: Transcriptional activation and DNAbinding by the erythroid factor GF-l/NF-El/Eryf 1. Genes Dev
41886,1990
28. Evans T, Felsenfeld G: Trans-activation of a globin promoter in non-erythroid cells. Mol Cell Biol11:843,1991
29. Fong TC, Emerson B M The erythroid-specific protein
cGATA-1 mediates distal enhancer activity through a specialized
@-globinTATA box. Genes Dev 6521,1992
30. Romeo P-H, Prandini M-H, Joulin V, Mignotte V, Prenant
M, Vainchenker W, Marguerie G, Uzan G: Megakaryocytic and
erythrocytic lineages share specific transcription factors. Nature
344:447,1990
31. Martin DIK, Zon LI, Mutter G, Orkin S H Expression of an
erythroid transcription factor in megakaryocytic and mast cell
lineages. Nature 344:444,1990
32. Zon LI, Gurish MF, Stevens RL, Mather C, Reynolds DS,
Austen KF, Orkin SH: GATA-binding transcription factors in mast
cells regulate the promoter of the mast cell carboxypeptidase A
gene. J Biol Chem 266:22948,1991
33. Ravid K, Doi T, Beeler DL, Kutter DJ, Rosenberg RD:
Transcriptionalregulation of the rat platelet factor 4 gene: Interaction between an enhancerhilencer domain and the GATA site.
Mol Cell Biolll:6116,1992
34. Wilson DB, Dorfman DM, Orkin SH: A non-erythroid
GATA-binding protein is required for function of the human
preproendothelin-1 promoter in endothelial cells. Mol Cell Biol
10:4854,1990
35. Evans T, Felsenfeld G: The erythroid-specific transcription
factor eryfl: A new finger protein. Cell 58:877,1989
36. Tsai SF, Martin DI, Zon LI, DAndrea AD, Wong GG,
Orkin S H Cloning of cDNA for the major DNA-binding protein of
the erythroid lineage through expression in mammalian cells.
Nature 339:446,1989
37. Yamamoto M, KO LJ, Leonard MW, Beug H, Orkin SH,
Engel JD: Activity and tissue-specificexpression of the transcription factor NF-E1 multigene family. Genes Dev 4:1650,1990
38. Kudla B, Caddick MX, Langdon T, Martinez-Rossi NM,
Bennett CF, Sibley S, Davies RW, Arst JHN. The regulatory gene
areA mediating nitrogen metabolite repression in Aspergillus
nidulans. Mutations affecting specificity of gene activation alter a
loop residue of a putative zinc finger. EMBO J 9:1355,1990
39. Spieth J, Shim YH, Lea K, Conrad R, Blumenthal T elt-1,
an embryonically expressed Caenorhabditis elegans gene homologous to the GATA transcription factor family. Mol Cell Biol
9:4651,1991
40. Zon LI, Mather C, Burgess S, Bolce ME, Harland RM,
Orkin SH: Expression of GATA-binding proteins during embryonic development in Xenopus laevis. Proc Natl Acad Sci USA
8810642,1991
41. Sposi NM, Zon LI, Care A, Valtieri M, Testa U, Gabbianelli
M, Mariani G, Bottero L, Mather C, Orkin SH, Peschle C:
Cycle-dependent initiation and lineage-dependent abrogation of
GATA-1 expression in pure differentiatinghematopoietic progenitors. Proc Natl Acad Sci USA 1992 (in press)
42. Crotta S, Nicolis S, Ronchi A, Ottolenghi S, Ruzzi L,
Shimada Y, Migliaccio AR, Migliaccio G: Progressive inactivation
of the expression of an erythroid transcriptional factor in GM- and
G-CSF-dependent myeloid cell lines. Nucleic Acids Res 18:6864,
1990
43. KO LJ, Yamamoto M, Leonard M W , George KM, Ting P,
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
GATA-BINDING TRANSCRIPTION FACTORS
Engel J D Murine and human T-lymphocyte GATA-3 factors
mediate transcription through a cis-regulatory element within the
human T-cell receptor 6 gene enhancer. Mol Cell Biol 11:2778,
1991
44. Ho I-C, Vorhees P, Marin N, Oakley BK, Tsai S-F, Orkin
SH, Leiden JM: Human GATA-3: A lineage-restricted transcription factor that regulates the expression of the T cell receptor a
gene. EMBO J 10:1187,1991
45. Joulin V, Bories D, Eleouet J-F, Labastie M-C, Chretien S,
Mattei M-G, Romeo P - H A T-cell specific TCR 6 DNA binding
protein is a member of the human GATA family. EMBO J 10:1809,
1991
46. Soriano P, Montgomery C, Geske R, Bradley A Targeted
disruption of the c-src proto-oncogene leads to osteopetrosis in
mice. Cell 64:693,1991
47. Capecchi MR: Altering the genome by homologousrecombination. Science 244:1288,1989
48. Wiles MV, Keller G: Multiple hematopoietic lineages develop from embryonic stem (ES) cells in culture. Development
111:259,1991
49. Doetschman T, Eistetter H, Katz M, Schmidt W, Kemler R:
The in vitro development of blastocyst-derivedembryonic stem cell
lines: Formation of visceral yolk sac, blood islands, and myocardium. J Embryo1 Exp Morpho1 8727,1985
50. Pevny L, Simon MC, Robertson E, Klein WH, Tsai S-F,
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D’Agati V, Orkin SH, Costantini F: Erythroid differentiation in
chimeric mice blocked by a targeted mutation in the gene for
transcription factor GATA-1. Nature 349:257,1991
51. Chapman VM, Stephenson DA, Mullins LJ, Keitz BT,
Disteche C, Orkin SH: Linkage of the erythroid transcription
factor gene (Gf-1) to the proximal region of the X chromosome of
mice. Genomics 9:309,1990
52. Simon MC, Pevny L, Wiles MV, Keller G, Costantini F,
Orkin SH: Rescue of erythroid development in gene targeted
GATA-1- mouse embryonic stem cells. Nature Genetics 1:92,1992
53. Begley CG, Aplan PD, Denning SM, Haynes BF, Waldmann
TA, Kirsch IR: The gene SCL is expressed during early hematopoiesis and encodes a differentiation-related DNA-binding motif.
Proc Natl Acad Sci USA 86:10128,1989
54. Chen Q, Cheng 1-T, Tsai L-H, Schneider N, Buchanan G,
Carroll A, Crist W, Ozanne B, Siciliano MJ, Baer R: The tal gene
undergoes chromosome translocation in T cell leukemia and
potentially encodes a helix-loop-helixprotein. EMBO J 9:415,1990
55. Weintraub H, Davis R, Tapscott S, Thayer M, Krause M,
Benezra R, Blackwell T, Turner D, Rupp R, Hollenberg S, Zhuang
Y, Lassar R: The myoD gene family: Nodal point during specification of the muscle cell lineage. Science 251:761,1991
56. Aplan PD, Begley CG, Bertness V, Nussmeier M, Ezquerra
A, Coligan J, Kirsch IR: The SCL gene is formed from a
transcriptionallycoinplex locus. Moi Cell Biol10:6426, 1990
From www.bloodjournal.org by guest on June 14, 2017. For personal use only.
1992 80: 575-581
GATA-binding transcription factors in hematopoietic cells
SH Orkin
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