A phosphorylation-and-ubiquitylation circuitry driving ATR activation and homologous recombination Journal Article
| Authors: | Dubois, J. C.; Yates, M.; Gaudreau-Lapierre, A.; Clément, G.; Cappadocia, L.; Gaudreau, L.; Zou, L.; Maréchal, A. |
| Article Title: | A phosphorylation-and-ubiquitylation circuitry driving ATR activation and homologous recombination |
| Abstract: | RPA-coated single-stranded DNA (RPA-ssDNA), a nucleoprotein structure induced by DNA damage, promotes ATR activation and homologous recombination (HR). RPA is hyper-phosphorylated and ubiquitylated after DNA damage. The ubiquitylation of RPA by PRP19 and RFWD3 facilitates ATR activation and HR, but how it is stimulated by DNA damage is still unclear. Here, we show that RFWD3 binds RPA constitutively, whereas PRP19 recognizes RPA after DNA damage. The recruitment of PRP19 by RPA depends on PIKK-mediated RPA phosphorylation and a positively charged pocket in PRP19. An RPA32 mutant lacking phosphorylation sites fails to recruit PRP19 and support RPA ubiquitylation. PRP19 mutants unable to bind RPA or lacking ubiquitin ligase activity also fail to support RPA ubiquitylation and HR. These results suggest that RPA phosphorylation enhances the recruitment of PRP19 to RPA-ssDNA and stimulates RPA ubiquitylation through a process requiring both PRP19 and RFWD3, thereby triggering a phosphorylation-ubiquitylation circuitry that promotes ATR activation and HR. © The Author(s) 2017. |
| Keywords: | signal transduction; genetics; dna replication; cytology; metabolism; dna damage; homologous recombination; dna repair; nuclear protein; ubiquitin protein ligase; cell line, tumor; hela cells; phosphorylation; nuclear proteins; gene expression regulation; chemistry; ubiquitination; tumor cell line; atm protein; dna repair enzymes; replication factor a; single stranded dna; dna, single-stranded; ubiquitin-protein ligases; atr protein, human; replication protein a; osteoblast; osteoblasts; dna ligase; hek293 cells; humans; human; hek293 cell line; hela cell line; ataxia telangiectasia mutated proteins; rna splicing factor; rna splicing factors; prpf19 protein, human; rfwd3 protein, human |
| Journal Title: | Nucleic Acids Research |
| Volume: | 45 |
| Issue: | 15 |
| ISSN: | 0305-1048 |
| Publisher: | Oxford University Press |
| Date Published: | 2017-09-06 |
| Start Page: | 8859 |
| End Page: | 8872 |
| Language: | English |
| DOI: | 10.1093/nar/gkx571 |
| PUBMED: | 28666352 |
| PROVIDER: | scopus |
| PMCID: | PMC5587784 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 2 November 2017 -- Source: Scopus |
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