A Yeast GSK-3 Kinase Mck1 Promotes Cdc6 Degradation to Inhibit DNA Re-Replication

Amy E. Ikui, Valentina Rossio, Lea Schroeder, Satoshi Yoshida

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Cdc6p is an essential component of the pre-replicative complex (pre-RC), which binds to DNA replication origins to promote initiation of DNA replication. Only once per cell cycle does DNA replication take place. After initiation, the pre-RC components are disassembled in order to prevent re-replication. It has been shown that the N-terminal region of Cdc6p is targeted for degradation after phosphorylation by Cyclin Dependent Kinase (CDK). Here we show that Mck1p, a yeast homologue of GSK-3 kinase, is also required for Cdc6 degradation through a distinct mechanism. Cdc6 is an unstable protein and is accumulated in the nucleus only during G1 and early S-phase in wild-type cells. In mck1 deletion cells, CDC6p is stabilized and accumulates in the nucleus even in late S phase and mitosis. Overexpression of Mck1p induces rapid Cdc6p degradation in a manner dependent on Threonine-368, a GSK-3 phosphorylation consensus site, and SCFCDC4. We show evidence that Mck1p-dependent degradation of Cdc6 is required for prevention of DNA re-replication. Loss of Mck1 activity results in synthetic lethality with other pre-RC mutants previously implicated in re-replication control, and these double mutant strains over-replicate DNA within a single cell cycle. These results suggest that a GSK3 family protein plays an unexpected role in preventing DNA over-replication through Cdc6 degradation in Saccharomyces cerevisiae. We propose that both CDK and Mck1 kinases are required for Cdc6 degradation to ensure a tight control of DNA replication.

Original languageEnglish
Article numbere1003099
JournalPLoS Genetics
Volume8
Issue number12
DOIs
Publication statusPublished - 2012 Dec 1
Externally publishedYes

Fingerprint

Glycogen Synthase Kinase 3
DNA Replication
yeast
phosphotransferases (kinases)
Phosphotransferases
Yeasts
DNA replication
yeasts
DNA
degradation
cyclin-dependent kinase
Cyclin-Dependent Kinases
S Phase
interphase
cell cycle
phosphorylation
Cell Cycle
Phosphorylation
replication origin
mutants

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Molecular Biology
  • Genetics
  • Genetics(clinical)
  • Cancer Research

Cite this

A Yeast GSK-3 Kinase Mck1 Promotes Cdc6 Degradation to Inhibit DNA Re-Replication. / Ikui, Amy E.; Rossio, Valentina; Schroeder, Lea; Yoshida, Satoshi.

In: PLoS Genetics, Vol. 8, No. 12, e1003099, 01.12.2012.

Research output: Contribution to journalArticle

Ikui, Amy E. ; Rossio, Valentina ; Schroeder, Lea ; Yoshida, Satoshi. / A Yeast GSK-3 Kinase Mck1 Promotes Cdc6 Degradation to Inhibit DNA Re-Replication. In: PLoS Genetics. 2012 ; Vol. 8, No. 12.
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