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

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Date
2010-05-01
Authors
Chee, Mark K
Haase, Steven B
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Abstract
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 SCFCdc4 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 (APCCdh1). 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 SCFCdc4 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. © 2010 Chee, Haase.
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Journal article
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https://hdl.handle.net/10161/4466
Published Version (Please cite this version)
10.1371/journal.pgen.1000935
Publication Info
Chee, Mark K; & Haase, Steven B (2010). B-Cyclin/CDKs regulate mitotic spindle assembly by phosphorylating kinesins-5 in budding yeast. PLoS Genetics, 6(5). pp. 35. 10.1371/journal.pgen.1000935. Retrieved from https://hdl.handle.net/10161/4466.
This is constructed from limited available data and may be imprecise. To cite this article, please review & use the official citation provided by the journal.
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Scholars@Duke

Haase

Steven B. Haase

Professor of Biology
Our group is broadly interested in understanding the biological clock mechanisms that control the timing of events during the cell division cycle. In 2008, the Haase group proposed a new model in which a complex network of sequentially activated transcription factors regulates the precise timing of gene expression during the cell-cycle, and functions as a robust time-keeping oscillator. Greater than a thousand genes are expressed at distinct phases of the cycle, and the control network itself
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