Rad3-dependent phosphorylation of the checkpoint clamp regulates repair-pathway choice

Mihoko Kai, Kanji Furuya, Francesca Paderi, Antony M. Carr, Teresa S F Wang

Research output: Contribution to journalArticle

Abstract

When replication forks collapse, Rad3 phosphorylates the checkpoint-clamp protein Rad9 in a manner that depends on Thr 225, a residue within the PCNA-like domain. The physiological function of Thr 225-dependent Rad9 phosphorylation, however, remains elusive. Here, we show that Thr 225-dependent Rad9 phosphorylation by Rad3 regulates DNA repair pathways. A rad9T225C mutant induces a translesion synthesis (TLS)-dependent high spontaneous mutation rate and a hyper-recombination phenotype. Consistent with this, Rad9 coprecipitates with the post-replication repair protein Mms2. This interaction is dependent on Rad9 Thr 225 and is enhanced by DNA damage. Genetic analyses indicate that Thr 225-dependent Rad9 phosphorylation prevents inappropriate Rhp51-dependent recombination, potentially by redirecting the repair through a Pli1-mediated sumoylation pathway into the error-free branch of the Rhp6 repair pathway. Our findings reveal a new mechanism by which phosphorylation of Rad9 at Thr 225 regulates the choice of repair pathways for maintaining genomic integrity during the cell cycle.

Original languageEnglish (US)
Pages (from-to)691-697
Number of pages7
JournalNature Cell Biology
Volume9
Issue number6
DOIs
StatePublished - Jun 2007
Externally publishedYes

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Phosphorylation
Genetic Recombination
Sumoylation
Proliferating Cell Nuclear Antigen
Mutation Rate
DNA Repair
DNA Damage
Cell Cycle
Phenotype
Proteins

ASJC Scopus subject areas

  • Cell Biology

Cite this

Rad3-dependent phosphorylation of the checkpoint clamp regulates repair-pathway choice. / Kai, Mihoko; Furuya, Kanji; Paderi, Francesca; Carr, Antony M.; Wang, Teresa S F.

In: Nature Cell Biology, Vol. 9, No. 6, 06.2007, p. 691-697.

Research output: Contribution to journalArticle

Kai, Mihoko ; Furuya, Kanji ; Paderi, Francesca ; Carr, Antony M. ; Wang, Teresa S F. / Rad3-dependent phosphorylation of the checkpoint clamp regulates repair-pathway choice. In: Nature Cell Biology. 2007 ; Vol. 9, No. 6. pp. 691-697.
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