Damage response of XRCC1 at sites of DNA single strand breaks is regulated by phosphorylation and ubiquitylation after degradation of poly(ADP-ribose)

Leizhen Wei, Satoshi Nakajima, Ching Lung Hsieh, Shinichiro Kanno, Mitsuko Masutani, Arthur S. Levine, Akira Yasui, Li Lan

Research output: Contribution to journalArticlepeer-review

44 Citations (Scopus)

Abstract

Single-strand breaks (SSBs) are the most common type of oxidative DNA damage and they are related to aging and many genetic diseases. The scaffold protein for repair of SSBs, XRCC1, accumulates at sites of poly(ADP-ribose) (pAR) synthesized by PARP, but it is retained at sites of SSBs after pAR degradation. How XRCC1 responds to SSBs after pAR degradation and how this affects repair progression are not well understood. We found that XRCC1 dissociates from pAR and is translocated to sites of SSBs dependent on its BRCTII domain and the function of PARG. In addition, phosphorylation of XRCC1 is also required for the proper dissociation kinetics of XRCC1 because (1) phosphorylation sites mutated in XRCC1 (X1 pm) cause retention of XRCC1 at sites of SSB for a longer time compared to wild type XRCC1; and (2) phosphorylation of XRCC1 is required for efficient polyubiquitylation of XRCC1. Interestingly, a mutant of XRCC1, LL360/361DD, which abolishes pAR binding, shows significant upregulation of ubiquitylation, indicating that pARylation of XRCC1 prevents the poly-ubiquitylation. We also found that the dynamics of the repair proteins DNA polymerase beta, PNK, APTX, PCNA and ligase I are regulated by domains of XRCC1. In summary, the dynamic damage response of XRCC1 is regulated in a manner that depends on modifications of polyADP-ribosylation, phosphorylation and ubiquitylation in live cells.

Original languageEnglish
Pages (from-to)4414-4423
Number of pages10
JournalJournal of cell science
Volume126
Issue number19
DOIs
Publication statusPublished - 2013

Keywords

  • Damage response
  • Phosphorylation
  • Polyadp-ribosylation
  • Single strand breaks
  • Ubiquitylation
  • Xrcc1

ASJC Scopus subject areas

  • Cell Biology

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