Rif1 controls DNA replication by directing Protein Phosphatase 1 to reverse Cdc7-mediated phosphorylation of the MCM complex

Shinichiro Hiraga, Gina M. Alvino, FuJung Chang, Huiyong Lian, Akila Sridhar, Takashi Kubota, Bonita J. Brewer, Michael Weinreich, M. K. Raghuraman, Anne D. Donaldson

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Abstract

Initiation of eukaryotic DNA replication requires phosphorylation of the MCM complex by Dbf4-dependent kinase (DDK), composed of Cdc7 kinase and its activator Dbf4. We report here that budding yeast Rif1 controls DNA replication genome-wide, and describe how Rif1 opposes DDK function by directing Protein Phosphatase 1 (PP1)-mediated dephosphorylation of the MCM complex. Deleting RIF1 partially compensates for the limited DDK activity in a cdc7-1 mutant strain by allowing increased, premature phosphorylation of Mcm4. PP1 interaction motifs within the Rif1 N-terminal domain are critical for its repressive effect on replication. We confirm that Rif1 interacts with PP1, and that PP1 prevents premature Mcm4 phosphorylation. Remarkably, our results suggest that replication repression by Rif1 is itself also DDK-regulated, through phosphorylation near the PP1-interacting motifs. Based on our findings, we propose that Rif1 is a novel PP1 substrate-targeting subunit that counteracts DDK-mediated phosphorylation during replication. Fission yeast and mammalian Rif1 proteins have also been implicated in regulating DNA replication. Since PP1 interaction sites are evolutionarily conserved within the Rif1 sequence, it is likely that replication control by Rif1 through PP1 is a conserved mechanism.
Original languageEnglish
Pages (from-to)372-383
Number of pages12
JournalGenes & Development
Volume28
Issue number4
DOIs
Publication statusPublished - 15 Feb 2014

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Protein Phosphatase 1
DNA Replication
Phosphorylation
Phosphotransferases
Saccharomycetales
Schizosaccharomyces
Genome

Keywords

  • DNA replication
  • protein phosphorylation
  • protein kinase
  • protein phosphatase
  • PP1

Cite this

Rif1 controls DNA replication by directing Protein Phosphatase 1 to reverse Cdc7-mediated phosphorylation of the MCM complex. / Hiraga, Shinichiro; Alvino, Gina M.; Chang, FuJung; Lian, Huiyong; Sridhar, Akila; Kubota, Takashi; Brewer, Bonita J.; Weinreich, Michael; Raghuraman, M. K.; Donaldson, Anne D.

In: Genes & Development, Vol. 28, No. 4, 15.02.2014, p. 372-383.

Research output: Contribution to journalArticle

Hiraga, Shinichiro ; Alvino, Gina M. ; Chang, FuJung ; Lian, Huiyong ; Sridhar, Akila ; Kubota, Takashi ; Brewer, Bonita J. ; Weinreich, Michael ; Raghuraman, M. K. ; Donaldson, Anne D. / Rif1 controls DNA replication by directing Protein Phosphatase 1 to reverse Cdc7-mediated phosphorylation of the MCM complex. In: Genes & Development. 2014 ; Vol. 28, No. 4. pp. 372-383.
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