Recovery from DNA damage checkpoint arrest by PP1-mediated inhibition of Chk1.
The G2 DNA damage checkpoint delays mitotic entry via the upregulation of Wee1 kinase and the downregulation of Cdc25 phosphatase by Chk1 kinase, and resultant inhibitory phosphorylation of Cdc2. While checkpoint activation is well understood, little is known about how the checkpoint is switched off to allow cell cycle re-entry. To identify proteins required for checkpoint release, we screened for genes in Schizosaccharomyces pombe that, when overexpressed, result in precocious mitotic entry in the presence of DNA damage. We show that overexpression of the type I protein phosphatase Dis2 sensitises S. pombe cells to DNA damage, causing aberrant mitoses. Dis2 abrogates Chk1 phosphorylation and activation in vivo, and dephosphorylates Chk1 and a phospho-S345 Chk1 peptide in vitro. dis2Delta cells have a prolonged chk1-dependent arrest and a compromised ability to downregulate Chk1 activity for checkpoint release. These effects are specific for the DNA damage checkpoint, because Dis2 has no effect on the chk1-independent response to stalled replication forks. We propose that inactivation of Chk1 by Dis2 allows mitotic entry following repair of DNA damage in the G2-phase.
den Elzen
Nicole R
NR
Trescowthick Research Laboratories, Peter MacCallum Cancer Centre, A'Beckett St., Melbourne, VIC, Australia.
O'Connell
Matthew J
MJ
D018797:Cell Cycle Proteins / Q000502:physiology; D000071877:Checkpoint Kinase 1; D064447:Checkpoint Kinase 2; D004249:DNA Damage; D004261:DNA Replication; D015536:Down-Regulation; D020869:Gene Expression Profiling; D008938:Mitosis; D009154:Mutation; D010749:Phosphoprotein Phosphatases / Q000096:biosynthesis / Q000502:physiology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000096:biosynthesis / Q000502:physiology*
D018797:Cell Cycle Proteins; D029702:Schizosaccharomyces pombe Proteins; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; D000071877:Checkpoint Kinase 1; C000605873:Chk1 protein, S pombe; C456840:rad3 protein, S pombe; C502092:Dis2 protein, S pombe; D010749:Phosphoprotein Phosphatases