A new XRCC1-containing complex and its role in cellular survival of methyl methanesulfonate treatment

Mol Cell Biol. 2004 Oct;24(19):8356-65. doi: 10.1128/MCB.24.19.8356-8365.2004.

Abstract

DNA single-strand break repair (SSBR) is important for maintaining genome stability and homeostasis. The current SSBR model derived from an in vitro-reconstituted reaction suggests that the SSBR complex mediated by X-ray repair cross-complementing protein 1 (XRCC1) is assembled sequentially at the site of damage. In this study, we provide biochemical data to demonstrate that two preformed XRCC1 protein complexes exist in cycling HeLa cells. One complex contains known enzymes that are important for SSBR, including DNA ligase 3 (DNL3), polynucleotide kinase 3'-phosphatase, and polymerase beta; the other is a new complex that contains DNL3 and the ataxia with oculomotor apraxia type 1 (AOA) gene product aprataxin. We report the characterization of the new XRCC1 complex. XRCC1 is phosphorylated in vivo and in vitro by CK2, and CK2 phosphorylation of XRCC1 on S518, T519, and T523 largely determines aprataxin binding to XRCC1 though its FHA domain. An acute loss of aprataxin by small interfering RNA renders HeLa cells sensitive to methyl methanesulfonate treatment by a mechanism of shortened half-life of XRCC1. Thus, aprataxin plays a role to maintain the steady-state protein level of XRCC1. Collectively, these data provide insights into the SSBR molecular machinery in the cell and point to the involvement of aprataxin in SSBR, thus linking SSBR to the neurological disease AOA.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acids / metabolism
  • Casein Kinase II
  • Cell Survival / physiology
  • DNA-Binding Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Methyl Methanesulfonate / metabolism*
  • Nuclear Proteins / metabolism
  • Phosphorylation
  • Protein Structure, Tertiary
  • Protein-Serine-Threonine Kinases / metabolism
  • X-ray Repair Cross Complementing Protein 1

Substances

  • APTX protein, human
  • Amino Acids
  • DNA-Binding Proteins
  • Nuclear Proteins
  • X-ray Repair Cross Complementing Protein 1
  • XRCC1 protein, human
  • Methyl Methanesulfonate
  • Casein Kinase II
  • Protein-Serine-Threonine Kinases