Mutations in ANP32A and ANP32C: influence on
function and potential implication in cancer
^
A thesis submitted for the degree of
Doctor of Philosophy (PhD)
in the subject of Biochemistry
by
Yuliia Yuzefovych, M.Sc. Biochemistry
September 2015
Hannover Medical School
International PhD program "Molecular Medicine"
in Hannover Biomedical Research School (HBRS)
Institute for Transfusion Medicine
http://d-nb.info/1081207582
TABLE OF CONTENTS
ABSTRACT
5
INTRODUCTION
7
1. ANP32 family: general characteristic and structure
7
2. ANP32A identification and functional overview
10
ANP32A in neurodegenerative disorders
11
ANP32A and PP2A inhibition
14
Role of ANP32A in cytoskeleton regulation and cellular transport
16
ANP32A and apoptosis
19
Nuclear functions of ANP32A
22
Other functions
27
3. Functions of ANP32B and ANP32E family members
32
4. Functions of ANP32C and ANP32D family members
40
5. Animal loss-of-function models
43
MATERIALS AND METHODS
46
UNPUBLISHED RESULTS 1
50
Mechanisms of the ANP32C/_Y140H oncogenic activity: Investigation of
the interaction with MORF4L1
50
1. Yeast two hybrid screening
50
2. Validation of the yeast two-hybrid screening results
50
PUBLICATION
53
Oncogenic acidic nuclear phosphoproteins ANP32C/D are novel clients of
heat shock protein 90
53
SUPLEMENTARY FIGURES
61
SUPPLEMENTARY METHODS
75
UNBUBLISHED RESULTS II
76
Characterization of the D146V ANP32A mutant
76
1. Effects on cellular proliferation
76
2. Stability assessment
76
3. Effects on the interaction with prominent ANP32A binding partners
78
4. Subcellular localization
81
5. Systemic effects on transcriptome
83
DISCUSSION
84
UNPUBLISHED RESULTS 1
84
3
ANP32C AND ANP32D ARE NOVEL CLIENTS OF HSP90
88
UNPUBLISHED RESULTS II
91
OUTLOOK
106
How to study ANP32 family?
106
What essential data are still missing?
107
Significance of the D146V mutation in ANP32A and the Y140H mutation in ANP32C
109
Can ANP32C be targeted in cancer with Hsp90 inhibitors?
110
BIBLIOGRAPHY
112
ACKNOWLEDGEMENTS
122
LIST OF PUBLICATIONS
123
CURRICULUM VITAE
124
DECLARATION
126
4