Regulation of Saccharomyces Rad53 checkpoint kinase during adaptation from DNA damage-induced G2/M arrest

Mol Cell. 2001 Feb;7(2):293-300. doi: 10.1016/s1097-2765(01)00177-0.

Abstract

Saccharomyces cells with one unrepaired double-strand break (DSB) adapt after checkpoint-mediated G2/M arrest. Adaptation is accompanied by loss of Rad53p checkpoint kinase activity and Chk1p phosphorylation. Rad53p kinase remains elevated in yku70delta and cdc5-ad cells that fail to adapt. Permanent G2/M arrest in cells with increased single-stranded DNA is suppressed by the rfa1-t11 mutation, but this RPA mutation does not suppress permanent arrest in cdc5-ad cells. Checkpoint kinase activation and inactivation can be followed in G2-arrested cells, but there is no kinase activation in G1-arrested cells. We conclude that activation of the checkpoint kinases in response to a single DNA break is cell cycle regulated and that adaptation is an active process by which these kinases are inactivated.

Publication types

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

MeSH terms

  • Adaptation, Biological / genetics*
  • Anaphase
  • Antigens, Nuclear*
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cell Cycle*
  • Checkpoint Kinase 1
  • Checkpoint Kinase 2
  • DNA Damage / genetics*
  • DNA Helicases*
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Enzyme Activation
  • Fungal Proteins / metabolism
  • G2 Phase
  • Gene Expression
  • Intracellular Signaling Peptides and Proteins
  • Kinetics
  • Ku Autoantigen
  • Mitosis
  • Mutation
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Phosphorylation
  • Protein Kinases / metabolism*
  • Protein Serine-Threonine Kinases*
  • RNA-Binding Proteins
  • Recombination, Genetic
  • Replication Protein A
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins*
  • Sequence Homology, Nucleic Acid

Substances

  • Antigens, Nuclear
  • CEF1 protein, S cerevisiae
  • Cell Cycle Proteins
  • DNA, Fungal
  • DNA-Binding Proteins
  • Fungal Proteins
  • Intracellular Signaling Peptides and Proteins
  • Nuclear Proteins
  • RNA-Binding Proteins
  • Replication Protein A
  • Saccharomyces cerevisiae Proteins
  • high affinity DNA-binding factor, S cerevisiae
  • Protein Kinases
  • Checkpoint Kinase 2
  • Checkpoint Kinase 1
  • MEC1 protein, S cerevisiae
  • Protein Serine-Threonine Kinases
  • RAD53 protein, S cerevisiae
  • DNA Helicases
  • XRCC5 protein, human
  • Xrcc6 protein, human
  • Ku Autoantigen