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.
| Original language | English |
|---|---|
| Journal | Journal of Cell Biology |
| Volume | 197 |
| Issue number | 2 |
| Pages (from-to) | 189-99 |
| Number of pages | 11 |
| ISSN | 0021-9525 |
| DOIs | |
| Publication status | Published - Apr 2012 |
Keywords
- BRCA1 Protein
- Carrier Proteins
- Cell Line, Tumor
- Cell Survival
- Chromatin
- DNA
- DNA Breaks, Double-Stranded
- DNA End-Joining Repair
- DNA-Binding Proteins
- HEK293 Cells
- HeLa Cells
- Homologous Recombination
- Humans
- Intracellular Signaling Peptides and Proteins
- Nuclear Proteins
- RNA Interference
- RNA, Small Interfering
- Ubiquitin
- Ubiquitin-Protein Ligases
- Ubiquitination
- Zinc Fingers