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Cytoplasmic Clb2 is required for timely inactivation of the mitotic inhibitor Swe1 and normal bud morphogenesis in Saccharomyces cerevisiae

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Abstract

Subcellular localization is an important determinant of substrate and functional specificity for cyclin–cyclin dependent kinase (CDK) complexes. This work addresses the cytoplasmic function of the budding yeast mitotic cyclin Clb2, which is mostly nuclear but is also present in the bulk cytoplasm and at the mother-bud neck. Clb2 contains two leucine-rich nuclear export signals (NESs)—one of which we newly describe here—that maintain its presence in the cytoplasm. Yeast strains bearing mutations in one or both of these NESs have elongated buds, indicative of a G2/M cell cycle delay. A small number of these cells exhibit a filamentous-like morphology under conditions that do not normally induce filamentous growth. These phenotypes are enhanced by deletion of the other three mitotic cyclins (CLB1,3,4) and are dependent on expression of Swe1, the yeast Cdk1 inhibitory kinase. Δclb1,3,4 Δbud3 cells, which fail to localize Clb2 to the bud neck, also exhibit a Swe1-dependent elongated bud phenotype. Our results support a model in which cytoplasmic Clb2-Cdk1 is required for timely inactivation of Swe1 at the G2/M transition and bud neck targeting of Clb2 contributes to the efficiency of this process. Cytoplasmic Clb2 may also be important for repression of filamentous growth.

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Acknowledgments

This work was supported by National Science Foundation grant MCB-0416738. C.N.B. and V.L. were supported by summer stipends from the Roberta Dey Staley and Karl A. Staley Fund for Cancer Research at Wellesley College. The authors thank Marie-Nöelle Simon, Anita Corbett, Doug Kellogg, David Pellman, and Fred Cross for providing strains and plasmids. They are also grateful to Caleb DeGrenier, Grace W. Wanjiku and Langdon Smythe for technical assistance and Jeff Hughes for consultation on statistical analysis.

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Authors and Affiliations

  1. Department of Biological Sciences, Wellesley College, 106 Central St., Wellesley, MA, 02481, USA

    Jennifer K. Hood-DeGrenier, Cortney N. Boulton & Victoria Lyo

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  1. Jennifer K. Hood-DeGrenier
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  2. Cortney N. Boulton
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  3. Victoria Lyo
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Correspondence to Jennifer K. Hood-DeGrenier.

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Communicated by P. Sunnerhagen.

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Hood-DeGrenier, J.K., Boulton, C.N. & Lyo, V. Cytoplasmic Clb2 is required for timely inactivation of the mitotic inhibitor Swe1 and normal bud morphogenesis in Saccharomyces cerevisiae . Curr Genet 51, 1–18 (2007). https://doi.org/10.1007/s00294-006-0102-1

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