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B-cyclin/CDKs regulate mitotic spindle assembly by phosphorylating kinesins-5 in budding yeast.

Publication ,  Journal Article
Chee, MK; Haase, SB
Published in: PLoS genetics
May 2010

Although it has been known for many years that B-cyclin/CDK complexes regulate the assembly of the mitotic spindle and entry into mitosis, the full complement of relevant CDK targets has not been identified. It has previously been shown in a variety of model systems that B-type cyclin/CDK complexes, kinesin-5 motors, and the SCF(Cdc4) ubiquitin ligase are required for the separation of spindle poles and assembly of a bipolar spindle. It has been suggested that, in budding yeast, B-type cyclin/CDK (Clb/Cdc28) complexes promote spindle pole separation by inhibiting the degradation of the kinesins-5 Kip1 and Cin8 by the anaphase-promoting complex (APC(Cdh1)). We have determined, however, that the Kip1 and Cin8 proteins are present at wild-type levels in the absence of Clb/Cdc28 kinase activity. Here, we show that Kip1 and Cin8 are in vitro targets of Clb2/Cdc28 and that the mutation of conserved CDK phosphorylation sites on Kip1 inhibits spindle pole separation without affecting the protein's in vivo localization or abundance. Mass spectrometry analysis confirms that two CDK sites in the tail domain of Kip1 are phosphorylated in vivo. In addition, we have determined that Sic1, a Clb/Cdc28-specific inhibitor, is the SCF(Cdc4) target that inhibits spindle pole separation in cells lacking functional Cdc4. Based on these findings, we propose that Clb/Cdc28 drives spindle pole separation by direct phosphorylation of kinesin-5 motors.

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Published In

PLoS genetics

DOI

EISSN

1553-7404

ISSN

1553-7390

Publication Date

May 2010

Volume

6

Issue

5

Start / End Page

e1000935

Related Subject Headings

  • Spindle Apparatus
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • Phosphorylation
  • Molecular Motor Proteins
  • Microtubule-Associated Proteins
  • Kinesins
  • Developmental Biology
  • Cyclin B
  • CDC28 Protein Kinase, S cerevisiae
 

Citation

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Chee, M. K., & Haase, S. B. (2010). B-cyclin/CDKs regulate mitotic spindle assembly by phosphorylating kinesins-5 in budding yeast. PLoS Genetics, 6(5), e1000935. https://doi.org/10.1371/journal.pgen.1000935
Chee, Mark K., and Steven B. Haase. “B-cyclin/CDKs regulate mitotic spindle assembly by phosphorylating kinesins-5 in budding yeast.PLoS Genetics 6, no. 5 (May 2010): e1000935. https://doi.org/10.1371/journal.pgen.1000935.
Chee, Mark K., and Steven B. Haase. “B-cyclin/CDKs regulate mitotic spindle assembly by phosphorylating kinesins-5 in budding yeast.PLoS Genetics, vol. 6, no. 5, May 2010, p. e1000935. Epmc, doi:10.1371/journal.pgen.1000935.

Published In

PLoS genetics

DOI

EISSN

1553-7404

ISSN

1553-7390

Publication Date

May 2010

Volume

6

Issue

5

Start / End Page

e1000935

Related Subject Headings

  • Spindle Apparatus
  • Saccharomyces cerevisiae Proteins
  • Saccharomyces cerevisiae
  • Phosphorylation
  • Molecular Motor Proteins
  • Microtubule-Associated Proteins
  • Kinesins
  • Developmental Biology
  • Cyclin B
  • CDC28 Protein Kinase, S cerevisiae