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PMID:19884341

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Citation

Bazzi, M, Mantiero, D, Trovesi, C, Lucchini, G and Longhese, MP (2010) Dephosphorylation of gamma H2A by Glc7/protein phosphatase 1 promotes recovery from inhibition of DNA replication. Mol. Cell. Biol. 30:131-45

Abstract

Replication fork stalling caused by deoxynucleotide depletion triggers Rad53 phosphorylation and subsequent checkpoint activation, which in turn play a crucial role in maintaining functional DNA replication forks. How cells regulate checkpoint deactivation after inhibition of DNA replication is poorly understood. Here, we show that the budding yeast protein phosphatase Glc7/protein phosphatase 1 (PP1) promotes disappearance of phosphorylated Rad53 and recovery from replication fork stalling caused by the deoxynucleoside triphosphate (dNTP) synthesis inhibitor hydroxyurea (HU). Glc7 is also required for recovery from a double-strand break-induced checkpoint, while it is dispensable for checkpoint inactivation during methylmethane sulfonate exposure, which instead requires the protein phosphatases Pph3, Ptc2, and Ptc3. Furthermore, Glc7 counteracts in vivo histone H2A phosphorylation on serine 129 (gamma H2A) and dephosphorylates gamma H2A in vitro. Finally, the replication recovery defects of HU-treated glc7 mutants are partially rescued by Rad53 inactivation or lack of gamma H2A formation, and the latter also counteracts hyperphosphorylated Rad53 accumulation. We therefore propose that Glc7 activity promotes recovery from replication fork stalling caused by dNTP depletion and that gamma H2A dephosphorylation is a critical Glc7 function in this process.

Links

PubMed PMC2798293 Online version:10.1128/MCB.01000-09

Keywords

Cell Cycle Proteins/metabolism; DNA Replication; DNA, Fungal/metabolism; Histones/metabolism; Hydroxyurea/pharmacology; Mutation; Phosphorylation; Protein Phosphatase 1/genetics; Protein Phosphatase 1/physiology; Protein-Serine-Threonine Kinases/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/metabolism; Saccharomyces cerevisiae Proteins/physiology

Significance

Annotations

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

YEAST:PP12

involved_in

GO:0000076: DNA replication checkpoint

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:PP12

involved_in

GO:0000076: DNA replication checkpoint

ECO:0000316: genetic interaction evidence used in manual assertion

SGD:S000000099
SGD:S000002633
SGD:S000006074

P

Seeded From UniProt

complete

YEAST:PP12

involved_in

GO:0000077: DNA damage checkpoint

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:PP12

involved_in

GO:0016576: histone dephosphorylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:PP12

involved_in

GO:0016576: histone dephosphorylation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:PP12

involved_in

GO:0031297: replication fork processing

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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