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Journal of Cell Science, Vol 112, Issue 14 2381-2390, Copyright © 1999 by Company of Biologists

JOURNAL ARTICLES

Functionally homologous DNA replication genes in fission and budding yeast

M Sanchez, A Calzada and A Bueno
Instituto de Microbiologia-Bioquimica/Centro de Investigacion del Cancer, Departamento de Microbiologia y Genetica, Edificio Departamental, Campus Miguel de Unamuno, CSIC/Universidad de Salamanca, Spain.

The cdc18(+) gene of the fission yeast Schizosaccharomyces pombe is involved in the initiation of DNA replication as well as in coupling the S phase to mitosis. In this work, we show that the Saccharomyces cerevisiae CDC6 gene complements cdc18-K46 ts and cdc18 deletion mutant S. pombe strains. The budding yeast gene suppresses both the initiation and the checkpoint defects associated with the lack of cdc18(+). The Cdc6 protein interacts in vivo with Cdc2 kinase complexes. Interestingly, Cdc6 is an in vitro substrate for Cdc13/Cdc2 and Cig1/Cdc2, but not for Cig2/Cdc2-associated kinases. Overexpression of Cdc6 in fission yeast induces multiple rounds of S-phase in the absence of mitosis and cell division. This CDC6-dependent continuous DNA synthesis phenotype is independent of the presence of a functional cdc18(+) gene product and, significantly, requires only Cig2/Cdc2-associated kinase activity. Finally, these S. pombe over-replicating cells do not require any protein synthesis other than that of Cdc6. Our data strongly suggest that CDC6 and cdc18(+) are functional homologues and also support the idea that controls restricting genome duplication diverge in fission and budding yeast.
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© The Company of Biologists Ltd 1999