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RESEARCH PAPER

Physical and functional interactions between polo kinase and the spindle pole component Cut12 regulate mitotic commitment in S. pombe

¹ Paterson Institute for Cancer Research, Manchester M20 4BX, UK , ² School of Biological Sciences, University of Manchester, Manchester M13 9PT, UK

Commitment to mitosis is regulated by a protein kinase complex called MPF. MPF is inhibited by Wee1-related kinases and activated by Cdc25 phosphatase. MPF activation further boosts Cdc25 and represses Wee1. This feedback control probably involves polo kinase. A dominant cut12.s11 mutation in the Schizosaccharomyces pombe spindle pole body (SPB) component Cut12 both suppresses the conditional lethal mitotic commitment defect of cdc25.22 and promotes premature association of the S. pombe polo kinase, Plo1, with the SPB. We now show that Cut12 associated with Plo1 in two hybrid and immunoprecipitation assays. Plo1 function was required for recognition of the mitotic SPB by the phospho-specific antibody MPM-2. In vivo MPM-2 staining and in vitro kinase assays established that the loss-of-function mutation, cut12.1, reduced mitotic activation of Plo1, whereas the gain-of-function mutation, cut12.s11, promoted higher levels of Plo1 activity than were normally seen in interphase. cut12.s11 could not promote mitotic commitment of cdc25.22 cells when Plo1 function was compromised. Expression of a constitutively active plo1 allele suppressed the mitotic commitment defect of cdc25.22. These data suggest that cut12.s11 suppresses cdc25.22 by promoting Plo1 activity. Furthermore, the delayed mitotic commitment of plo1.ts2 cells suggests that Plo1 is an integral part of the core controls that modulate MPF activation in S. pombe.

[Keywords: S. pombe; polo kinase; cut12; mitosis; MPF; centrosome; SPB]

Received November 28, 2002; revised version accepted April 15, 2003.

³ These authors contributed equally to this work.

⁴ Present address: Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, 7-3-1 Hongo, Tokyo, Japan

⁵ E-MAIL ihagan{at}picr.man.ac.uk; FAX 44-161-446-3109.

Article and publication are at http://www.genesdev.org/cgi/doi/10.1101/gad.256003.

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