Src family kinases promote silencing of ATR-Chk1 signaling in termination of DNA damage checkpoint

J Biol Chem. 2014 May 2;289(18):12313-29. doi: 10.1074/jbc.M113.533752. Epub 2014 Mar 14.

Abstract

The DNA damage checkpoint arrests cell cycle progression to allow time for repair. Once DNA repair is completed, checkpoint signaling is terminated. Currently little is known about the mechanism by which checkpoint signaling is terminated, and the disappearance of DNA lesions is considered to induce the end of checkpoint signaling; however, here we show that the termination of checkpoint signaling is an active process promoted by Src family tyrosine kinases. Inhibition of Src activity delays recovery from the G2 phase DNA damage checkpoint following DNA repair. Src activity is required for the termination of checkpoint signaling, and inhibition of Src activity induces persistent activation of ataxia telangiectasia mutated (ATM)- and Rad3-related (ATR) and Chk1 kinases. Src-dependent nuclear protein tyrosine phosphorylation and v-Src expression suppress the ATR-mediated Chk1 and Rad17 phosphorylation induced by DNA double strand breaks or DNA replication stress. Thus, Src family kinases promote checkpoint recovery through termination of ATR- and Chk1-dependent G2 DNA damage checkpoint. These results suggest a model according to which Src family kinases send a termination signal between the completion of DNA repair and the initiation of checkpoint termination.

Keywords: Cell Cycle; Checkpoint Control; Checkpoint Recovery; DNA Damage; DNA Damage Response; Src.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ataxia Telangiectasia Mutated Proteins / genetics
  • Ataxia Telangiectasia Mutated Proteins / metabolism
  • Cell Cycle Proteins / metabolism
  • Checkpoint Kinase 1
  • DNA Damage*
  • DNA Repair
  • Down-Regulation / drug effects
  • Doxorubicin / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Flow Cytometry
  • G2 Phase Cell Cycle Checkpoints / drug effects
  • G2 Phase Cell Cycle Checkpoints / genetics
  • HeLa Cells
  • Humans
  • Indoles / pharmacology
  • Microscopy, Fluorescence
  • Nuclear Proteins / metabolism
  • Phosphorylation / drug effects
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • RNA Interference
  • Signal Transduction / drug effects
  • Signal Transduction / genetics
  • Sulfonamides / pharmacology
  • Topoisomerase II Inhibitors / pharmacology
  • Tyrosine / metabolism
  • src-Family Kinases / antagonists & inhibitors
  • src-Family Kinases / genetics
  • src-Family Kinases / metabolism*

Substances

  • Cell Cycle Proteins
  • Indoles
  • Nuclear Proteins
  • Rad17 protein, human
  • SU 6656
  • Sulfonamides
  • Topoisomerase II Inhibitors
  • Tyrosine
  • Doxorubicin
  • Protein Kinases
  • src-Family Kinases
  • ATR protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • CHEK1 protein, human
  • Checkpoint Kinase 1