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First published online 9 January 2007
doi: 10.1242/jcs.03342

Journal of Cell Science 120, 435-446 (2007)
Published by The Company of Biologists 2007
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Research Article

Phosphorylation of Spc110p by Cdc28p-Clb5p kinase contributes to correct spindle morphogenesis in S. cerevisiae

Stephen M. Huisman, Monique F. M. A. Smeets* and Marisa Segal{ddagger}

Department of Genetics, University of Cambridge, Downing Street, Cambridge, CB2 3EH, UK

{ddagger} Author for correspondence (e-mail: ms433{at}cam.ac.uk)

Accepted 14 November 2006

Spindle morphogenesis is regulated by cyclin-dependent kinases and monitored by checkpoint pathways to accurately coordinate chromosomal segregation with other events in the cell cycle. We have previously dissected the contribution of individual B-type cyclins to spindle morphogenesis in Saccharomyces cerevisiae. We showed that the S-phase cyclin Clb5p is required for coupling spindle assembly and orientation. Loss of Clb5p-dependent kinase abolishes intrinsic asymmetry between the spindle poles resulting in lethal translocation of the spindle into the bud with high penetrance in diploid cells. This phenotype was exploited in a screen for high dosage suppressors that yielded spc110{Delta}13, encoding a truncation of the spindle pole body component Spc110p (the intranuclear receptor for the {gamma}-tubulin complex). We found that Clb5p-GFP was localised to the spindle poles and intranuclear microtubules and that Clb5p-dependent kinase promoted cell cycle dependent phosphorylation of Spc110p contributing to spindle integrity. Two cyclin-dependent kinase consensus sites were required for this phosphorylation and were critical for the activity of spc110{Delta}13 as a suppressor. Together, our results point to the function of cyclin-dependent kinase phosphorylation of Spc110p and provide, in addition, support to a model for Clb5p control of spindle polarity at the level of astral microtubule organisation.

Key words: Spindle, Cell cycle, Cyclin-dependent kinase, Microtubule organisation


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[Abstract] [Full Text] [PDF]


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