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PMID:11313465

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Citation

Tanaka, K, Boddy, MN, Chen, XB, McGowan, CH and Russell, P (2001) Threonine-11, phosphorylated by Rad3 and atm in vitro, is required for activation of fission yeast checkpoint kinase Cds1. Mol. Cell. Biol. 21:3398-404

Abstract

Fission yeast Cds1 is phosphorylated and activated when DNA replication is interrupted by nucleotide starvation or DNA damage. Cds1 enforces the S-M checkpoint that couples mitosis (M) to the completion of DNA synthesis (S). Cds1 also controls replicational stress tolerance mechanisms. Cds1 is regulated by a group of proteins that includes Rad3, a kinase related to human checkpoint kinase ATM (ataxia telangiectasia mutated). ATM phosphorylates serine or threonine followed by glutamine (SQ or TQ). Here we show that in vitro, Rad3 and ATM phosphorylate the N-terminal domain of Cds1 at the motif T(11)Q(12). Substitution of threonine-11 with alanine (T11A) abolished Cds1 activation that occurs when DNA replication is inhibited by hydroxyurea (HU) treatment. The cds1-T11A mutant was profoundly sensitive to HU, although not quite as sensitive as a cds1(-) null mutant. Cds1(T11A) was unable to enforce the S-M checkpoint. These results strongly suggest that Rad3-dependent phosphorylation of Cds1 at threonine-11 is required for Cds1 activation and function.

Links

PubMed PMC100261 Online version:10.1128/MCB.21.10.3398-3404.2001

Keywords

Adenosine Triphosphatases/metabolism; Checkpoint Kinase 2; DNA Helicases/metabolism; Enzyme Activation; Phosphorylation; Protein Kinases/metabolism; Protein-Serine-Threonine Kinases; Saccharomyces cerevisiae Proteins; Schizosaccharomyces/enzymology; Schizosaccharomyces pombe Proteins; Threonine

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

SCHPO:RAD3

involved_in

GO:0033314: mitotic DNA replication checkpoint signaling

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

SCHPO:RAD3

enables

GO:0004674: protein serine/threonine kinase activity

ECO:0000314: direct assay evidence used in manual assertion

F

  • has_input:(PomBase:SPCC18B5.11c)
  • part_of:(GO:0033314)

Seeded From UniProt

complete

SCHPO:CDS1

involved_in

GO:0033314: mitotic DNA replication checkpoint signaling

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

SCHPO:CDS1

enables

GO:0004674: protein serine/threonine kinase activity

ECO:0000314: direct assay evidence used in manual assertion

F

  • has_input:(PomBase:SPCC18B5.03)
  • part_of:(GO:0033314)

Seeded From UniProt

complete

SCHPO:RAD3

involved_in

GO:0033314: mitotic DNA replication checkpoint

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

SCHPO:RAD3

enables

GO:0004674: protein serine/threonine kinase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

Notes

See also

References

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