DOI: 10.1073/pnas.221467598. Redundant control of rereplication in fission yeast

Redundant control of rereplication in fission yeast

10.1073/pnas.221467598

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The initiation of DNA replication at replication origins in eukaryotic cells is tightly controlled to ensure that the genome is duplicated only once each cell cycle. We present evidence that in fission yeast, independent regulation of two essential components of the initiation complex, Cdc18 and Cdt1, contributes to the prevention of reinitiation of DNA replication. Cdc18 is negatively controlled by cyclin-dependent kinase (CDK) phosphorylation, but low level expression of a mutant form of Cdc18 lacking CDK phosphorylation sites (Cdc18 CDK ) is not sufficient to induce rereplication. Similar to Cdc18, Cdt1 is expressed periodically in the cell cycle, accumulating in the nucleus in G 1 and declining in G 2 . When Cdt1 is expressed constitutively from an ectopic promoter, it accumulates in the nucleus throughout the cell cycle but does not promote reinitiation. However, constitutive expression of Cdt1, together with Cdc18 CDK , is sufficient to induce extra rounds of DNA replication in the absence of mitosis. Significantly greater levels of rereplication can be induced by coexpression of Cdc18 CDK and a Cdt1 mutant lacking a conserved C-terminal motif. In contrast, uncontrolled DNA replication does not occur when either mutant protein is expressed in the absence of the other. Constitutive expression of wild-type or mutant Cdt1 also leads to an increase in the levels of Cdc18 CDK , possibly as a result of increased protein stability. Our data are consistent with the hypothesis that control of rereplication depends on a redundant mechanism in which negative regulation of Cdt1 functions in parallel with the negative regulation of Cdc18.

Bibliography

Gopalakrishnan, V., Simancek, P., Houchens, C., Snaith, H. A., Frattini, M. G., Sazer, S., & Kelly, T. J. (2001). Redundant control of rereplication in fission yeast. Proceedings of the National Academy of Sciences, 98(23), 13114â13119.

Authors 7

Vidya Gopalakrishnan (first)
Pamela Simancek (additional)
Christopher Houchens (additional)
Hilary A. Snaith (additional)
Mark G. Frattini (additional)
Shelley Sazer (additional)
Thomas J. Kelly (additional)

References 36 Referenced 44

Dates

Type	When
Created	23 years ago (July 26, 2002, 10:32 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 8:34 a.m.)
Indexed	2 months, 1 week ago (June 10, 2025, 2:44 p.m.)
Issued	23 years, 10 months ago (Oct. 23, 2001)
Published	23 years, 10 months ago (Oct. 23, 2001)
Published Online	23 years, 10 months ago (Oct. 23, 2001)
Published Print	23 years, 9 months ago (Nov. 6, 2001)

Funders 0

None

BibTeX

@article{Gopalakrishnan_2001, title={Redundant control of rereplication in fission yeast}, volume={98}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.221467598}, DOI={10.1073/pnas.221467598}, number={23}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Gopalakrishnan, Vidya and Simancek, Pamela and Houchens, Christopher and Snaith, Hilary A. and Frattini, Mark G. and Sazer, Shelley and Kelly, Thomas J.}, year={2001}, month=oct, pages={13114–13119} }

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  "abstract": "<jats:p>\n            The initiation of DNA replication at replication origins in eukaryotic cells is tightly controlled to ensure that the genome is duplicated only once each cell cycle. We present evidence that in fission yeast, independent regulation of two essential components of the initiation complex, Cdc18 and Cdt1, contributes to the prevention of reinitiation of DNA replication. Cdc18 is negatively controlled by cyclin-dependent kinase (CDK) phosphorylation, but low level expression of a mutant form of Cdc18 lacking CDK phosphorylation sites (Cdc18\n            <jats:sup>CDK</jats:sup>\n            ) is not sufficient to induce rereplication. Similar to Cdc18, Cdt1 is expressed periodically in the cell cycle, accumulating in the nucleus in G\n            <jats:sub>1</jats:sub>\n            and declining in G\n            <jats:sub>2</jats:sub>\n            . When Cdt1 is expressed constitutively from an ectopic promoter, it accumulates in the nucleus throughout the cell cycle but does not promote reinitiation. However, constitutive expression of Cdt1, together with Cdc18\n            <jats:sup>CDK</jats:sup>\n            , is sufficient to induce extra rounds of DNA replication in the absence of mitosis. Significantly greater levels of rereplication can be induced by coexpression of Cdc18\n            <jats:sup>CDK</jats:sup>\n            and a Cdt1 mutant lacking a conserved C-terminal motif. In contrast, uncontrolled DNA replication does not occur when either mutant protein is expressed in the absence of the other. Constitutive expression of wild-type or mutant Cdt1 also leads to an increase in the levels of Cdc18\n            <jats:sup>CDK</jats:sup>\n            , possibly as a result of increased protein stability. Our data are consistent with the hypothesis that control of rereplication depends on a redundant mechanism in which negative regulation of Cdt1 functions in parallel with the negative regulation of Cdc18.\n          </jats:p>",
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