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. Deltaclb1,3,4 Deltabud3 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.