Regulation of hypoxia-inducible factors by small ubiquitin-like modifiers
Hagen, M. van
Citation
Hagen, M. van. (2010, May 26). Regulation of hypoxia-inducible factors by small ubiquitin- like modifiers. Retrieved from https://hdl.handle.net/1887/15552
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Regulation of Hypoxia-Inducible Factors by Small Ubiquitin-like Modifiers
Martijn van Hagen
Regulation of Hypoxia-Inducible Factors by Small Ubiquitin-like Modifiers
Proefschrift
ter verkrijging van
de graad van Doctor aan de Universiteit Leiden,
op gezag van Rector Magnificus prof. mr. P.F. van der Heijden, volgens besluit van het College voor Promoties
te verdedigen op woensdag 26 mei 2010 klokke 13.45 uur
door
Martijn van Hagen
geboren te Delft
in 1979
Promotiecommissie
Promotor: Prof. Dr. H.J. Tanke Co-promotor: Dr. A.C.O. Vertegaal Overige leden: Prof. Dr. P. ten Dijke
Prof. Dr. L.H. Mullenders
Dr. M.A. Vooijs (UMC Utrecht en UMC Maastricht)
The studies described in this thesis were performed at the department of Molecular and Cellular Biology, Leiden University Medical Center.
This thesis was printed by Gildeprint Drukkerijen, Enschede.
Table of contents
Outline of the thesis 7
List of abbreviations used in the thesis 9
General introduction 15
Chapter 1: Introduction 1: Small Ubiquitin-like Modifiers M. van Hagen and A.C.O. Vertegaal.
Small Ubiquitin-like Modifiers and other Ubiquitin-like Proteins.
Vidal, CJ (ed.), Protein reviews series: Post-translational modifications in health and disease. (2010) Springer Business and media.
17
Chapter 2: Introduction 2: Hypoxia-Inducible Factors 47
Chapter 3: Inhibition of Hypoxia-Inducible Factor-1αααα by conjugation of Small Ubiquitin-like Modifiers
M. van Hagen, S.S. Abolvardi, J.V. Olsen, C.I. Balog, G. Roukens, D. Baker, P. Hensbergen and A.C.O. Vertegaal.
Submitted for publication.
61
Chapter 4: The ubiquitin-proteasome system is a key component of the SUMO-2/3 cycle J. Schimmel, K.M. Larsen, I. Matic, M. van Hagen, J. Cox, M. Mann,
J.S. Andersen and A.C.O. Vertegaal Mol. Cell Proteomics (2008), 7, 2107-2122.
89
Chapter 5: RNF4 and VHL regulate the proteasomal degradation of SUMO-conjugated Hypoxia-Inducible Factor-2αααα
M. van Hagen, R.M. Overmeer, S.S. Abolvardi and A.C.O. Vertegaal.
Nucleic Acids Res. (2010), 38, 1922-1931.
107
Chapter 6: In vivo identification of human Small Ubiquitin-like Modifier polymerization sites by high accuracy mass spectrometry and an in vitro to in vivo strategy I. Matic, M. van Hagen, J. Schimmel, B. Macek, S.C. Ogg, M.H. Tatham, R.T. Hay, A.I. Lamond, M. Mann and A.C.O. Vertegaal
Mol. Cell Proteomics (2008), 7, 132-144.
123
Chapter 7: Summary and discussion 139
Chapter 8: Nederlandse samenvatting 149
Curriculum Vitae 155
List of publications 157
7
Outline of the thesis
The topic of this thesis is the molecular regulation of Hypoxia-Inducible Factors (HIFs) by sumoylation. Hypoxia-Inducible Factors are transcriptional regulators that mediate cell survival during reduced oxygen conditions (hypoxia). HIFs are extensively controlled by post-translational modifications. The aim of this thesis was to obtain novel insight in the post- translational modification of HIFs by sumoylation.
Chapters 1 and 2 are introductions on sumoylation and Hypoxia-Inducible Factors, respectively. A summary of current knowledge is provided based on published literature on both systems.
Chapter 3 describes the regulation of HIF-1α transcriptional activity by sumoylation via a novel SUMO acceptor site.
Chapter 4 describes crosstalk between sumoylation and ubiquitination, two similar but distinct post-translational modifications that act in concert to regulate the proteasomal degradation of a subset of protein targets.
Chapter 5 shows that sumoylation of HIF-2α marks it for proteasomal degradation during hypoxia, facilitated by E3 ubiquitin ligases RNF4 and VHL.
Chapter 6 presents data on the formation of SUMO chains made up of a mix of SUMO-1 and SUMO-2/3 and we show that HIF-1α is a target for SUMO chains in vitro.
Chapter 7 provides a summary and overall discussion of the thesis.
8
9
List of abbreviations used in the thesis
Aos1 Activation of Smt3p
ATG8 Autophagy-related protein 8 ATG12 Autophagy-related protein 12 bHLH basic helix-loop-helix
CBP cAMP response element binding (CREB)-binding protein CK2 Casein kinase 2
C-TAD C-terminal transactivation domain E1 SUMO activating enzyme
E2 SUMO protein carrier protein
E3 SUMO ligase
EPO Erythropoietin
EPAS Endothelial PAS domain protein ES Embryonic stem (cell)
FAT10 Ubiquitin-like protein FAT10 FIH Factor inhibiting HIF
FUBI Ubiquitin-like protein FUBI GMP1 GAP-modifying protein 1 HDAC Histone deacetylase
HECT Homologous to the E6-AP Carboxyl Terminus HIF Hypoxia-inducible factor
HSP90 Heat-shock protein 90
HUB1 Ubiquitin-like modifier HUB1 IκBα NFκB inhibitor α
iNOS Inducible nitric oxide synthase IR Internal repeats
ISG15 Interferon-stimulated gene 15 MAPK Mitogen-activated protein kinase Mdm2 Mouse double minute 2
MDR Multidrug resistance (gene)
NDSM Negatively charged amino acid-dependent sumoylation motif
NEDD8 Neural precursor cell expressed developmentally down-regulated protein 8
10 NFκB Nuclear factor κB
NO Nitric oxide NPC Nuclear pore complex
N-TAD N-terminal transactivation domain ODD Oxygen-dependent degradation (domain)
PAS Per-ARNT-SIM
Pc2 Polycomb protein 2
PCNA Proliferating cell nuclear antigen
PDSM Phosphorylation-dependent sumoylation motif PHD Proline hydroxylase
PIAS Protein inhibitor of activated STAT PIC1 PML-interacting clone 1
PML Promyelocytic leukemia protein PUP Prokaryotic ubiquitin-like protein RACK1 Receptor of activated protein C kinase 1 RanBP Ran-binding protein
RanGAP Ran GTPase-activating protein RING Really interesting new gene
RNF4 RING finger protein 4
RSUME RWD domain-containing sumoylation enhancer
RWD RING finger- and WD repeat-containing proteins and yeast DEAD-like helicases
SAE SUMO activating enzyme SENP SUMO protease
SILAC Stable-isotope labeling by amino acids in cell culture SIM SUMO interaction motif
Siz SAP and Miz-finger domain-containing protein 1 SMT3 Suppressor of mif two 3
STAT Signal transducer and activator of transcription SUMO Small ubiquitin-like modifier
TAF TBP-associated factor TBP TATA box binding protein TDG Thymine-DNA glycosylase
11 Uba2 Ubiquitin-activating enzyme E1-like Ubc9 Ubiquitin carrier protein 9
UBL1 Ubiquitin-like protein 1 UFM1 Ubiquitin-fold modifier 1 Ulp Ubiquitin-like-specific protease URM1 Ubiquitin-related modifier 1
VDU2 VHL-interacting deubiquitinating (enzyme) 2 VEGF Vascular endothelial growth factor
VHL Von Hippel Lindau
12
