HERC2 coordinates ubiquitin-dependent assembly of DNA repair factors on damaged chromosomes

Simon Bekker-Jensen; Jannie Rendtlew Danielsen; Kasper Fugger; Irina Gromova; Annika Nerstedt; Claudia Lukas; Jiri Bartek; Jiri Lukas; Niels Mailand

doi:10.1038/ncb2008

HERC2 coordinates ubiquitin-dependent assembly of DNA repair factors on damaged chromosomes

Nat Cell Biol. 2010 Jan;12(1):80-6; sup pp 1-12. doi: 10.1038/ncb2008. Epub 2009 Dec 20.

Authors

Simon Bekker-Jensen¹, Jannie Rendtlew Danielsen, Kasper Fugger, Irina Gromova, Annika Nerstedt, Claudia Lukas, Jiri Bartek, Jiri Lukas, Niels Mailand

Affiliation

¹ Centre for Genotoxic Stress Research, Department of Proteomics in Cancer, Institute of Cancer Biology, Danish Cancer Society, Copenhagen, Denmark.

PMID: 20023648
DOI: 10.1038/ncb2008

Abstract

Regulatory ubiquitylation is emerging as an important mechanism to protect genome integrity in cells exposed to DNA damage. However, the spectrum of known ubiquitin regulators of the DNA damage response (DDR) is limited and their functional interplay is poorly understood. Here, we identify HERC2 as a factor that regulates ubiquitin-dependent retention of repair proteins on damaged chromosomes. In response to ionising radiation (IR), HERC2 forms a complex with RNF8, a ubiquitin ligase involved in the DDR. The HERC2-RNF8 interaction requires IR-inducible phosphorylation of HERC2 at Thr 4827, which in turn binds to the forkhead-associated (FHA) domain of RNF8. Mechanistically, we provide evidence that HERC2 facilitates assembly of the ubiquitin-conjugating enzyme Ubc13 with RNF8, thereby promoting DNA damage-induced formation of Lys 63-linked ubiquitin chains. We also show that HERC2 interacts with, and maintains the levels of, RNF168, another ubiquitin ligase operating downstream of RNF8 (Refs 7, 8). Consequently, knockdown of HERC2 abrogates ubiquitin-dependent retention of repair factors such as 53BP1, RAP80 and BRCA1. Together with the increased radiosensitivity of HERC2-depleted cells, these results uncover a regulatory layer in the orchestration of protein interactions on damaged chromosomes and they underscore the role of ubiquitin-mediated signalling in genome maintenance.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

BRCA1 Protein / genetics
BRCA1 Protein / metabolism
Bone Neoplasms / genetics
Bone Neoplasms / metabolism
Carrier Proteins / genetics
Carrier Proteins / metabolism
Cells, Cultured
Chromosomes, Human / radiation effects*
DNA Damage / physiology*
DNA Repair*
DNA-Binding Proteins / metabolism*
Guanine Nucleotide Exchange Factors / antagonists & inhibitors
Guanine Nucleotide Exchange Factors / physiology*
Histone Chaperones
Humans
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Kidney / cytology
Kidney / metabolism
Lysine / genetics
Lysine / metabolism
Mutation / genetics
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Osteosarcoma / genetics
Osteosarcoma / metabolism
Phosphorylation
RNA, Small Interfering / pharmacology
Tumor Suppressor p53-Binding Protein 1
Ubiquitin / metabolism*
Ubiquitin-Protein Ligases / metabolism*
Ubiquitination
Ultraviolet Rays

Substances

BRCA1 Protein
BRCA1 protein, human
Carrier Proteins
DNA-Binding Proteins
Guanine Nucleotide Exchange Factors
Histone Chaperones
Intracellular Signaling Peptides and Proteins
Nuclear Proteins
RNA, Small Interfering
RNF8 protein, human
TP53BP1 protein, human
Tumor Suppressor p53-Binding Protein 1
UIMC1 protein, human
Ubiquitin
HERC2 protein, human
Ubiquitin-Protein Ligases
Lysine