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PMID: 22492721 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Human RNF169 is a negative regulator of the ubiquitin-dependent response to DNA double-strand breaks.

The Journal of cell biology ·Vol. 197 ·No. 2 ·2012-04-16 ·Pages 189-99

Poulsen M, Lukas C, Lukas J, Bekker-Jensen S, Mailand N

Abstract

Nonproteolytic ubiquitylation of chromatin surrounding deoxyribonucleic acid double-strand breaks (DSBs), mediated by the RNF8/RNF168 ubiquitin ligases, plays a key role in recruiting repair factors, including 53BP1 and BRCA1, to reestablish genome integrity. In this paper, we show that human RNF169, an uncharacterized E3 ubiquitin ligase paralogous to RNF168, accumulated in DSB repair foci through recognition of RNF168-catalyzed ubiquitylation products by its motif interacting with ubiquitin domain. Unexpectedly, RNF169 was dispensable for chromatin ubiquitylation and ubiquitin-dependent accumulation of repair factors at DSB sites. Instead, RNF169 functionally competed with 53BP1 and RAP80-BRCA1 for association with RNF168-modified chromatin independent of its catalytic activity, limiting the magnitude of their recruitment to DSB sites. By delaying accumulation of 53BP1 and RAP80 at damaged chromatin, RNF169 stimulated homologous recombination and restrained nonhomologous end joining, affecting cell survival after DSB infliction. Our results show that RNF169 functions in a noncanonical fashion to harness RNF168-mediated protein recruitment to DSB-containing chromatin, thereby contributing to regulation of DSB repair pathway utilization.

MeSH Terms
BRCA1 Protein/metabolism Carrier Proteins/metabolism Cell Line, Tumor Cell Survival/genetics Chromatin/metabolism DNA/genetics,metabolism DNA Breaks, Double-Stranded DNA End-Joining Repair DNA-Binding Proteins/genetics HEK293 Cells HeLa Cells Histone Chaperones Homologous Recombination Humans Intracellular Signaling Peptides and Proteins/metabolism Nuclear Proteins/metabolism RNA Interference RNA, Small Interfering Tumor Suppressor p53-Binding Protein 1 Ubiquitin/metabolism Ubiquitin-Protein Ligases/genetics,metabolism Ubiquitination Zinc Fingers/genetics
Chemicals
BRCA1 Protein BRCA1 protein, human Carrier Proteins Chromatin DNA-Binding Proteins 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 DNA RNF168 protein, human RNF169 protein, human Ubiquitin-Protein Ligases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Poulsen Maria
Ubiquitin Signaling Group, Department of Disease Biology, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, DK-2200 Copenhagen, Denmark.
Lukas Claudia
Lukas Jiri
Bekker-Jensen Simon
Mailand Niels
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201109100
Published
2012-04-16
Epub
2012-00-09
Pages
189-99
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC3328375
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.201109100 PMC Full Text View journal details
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