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Krisada Sakchaisri*,,
Satoshi Asano*,
Li-Rong Yu,
Mark J. Shulewitz,
Chong J. Park*,
Jung-Eun Park*,
Young-Wook Cho*,
Timothy D. Veenstra,
Jeremy Thorner, and
Kyung S. Lee*,¶
*Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892; Mass Spectrometry Center, National Cancer Institute–Frederick, Frederick, MD 21702; Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720; and Department of Pharmacology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
Received for publication December 22, 2003.
Abstract:
In eukaryotes, cyclin B-bound cyclin-dependent protein kinase1 promotes mitotic entry but is held in check, in part, by Wee1protein kinase. Timely mitotic entry in budding yeast requiresinactivation of Swe1 (Wee1 ortholog). Perturbations of the septincollar at the bud neck lead to Swe1 stabilization, delayingthe G2/M transition. Swe1 is recruited to the neck and hyperphosphorylatedbefore ubiquitin-mediated degradation. Hsl1 kinase (Nim1 ortholog),a negative regulator of Wee1, is required for efficient Swe1localization at the neck but seems not to phosphorylate Swe1.Here, we show that two other kinases targeted sequentially tothe neck, Cla4/PAK and Cdc5/Polo, are responsible for stepwisephosphorylation and down-regulation of Swe1. This mechanismlinks assembly of a cellular structure to passage into mitosis.
¶ To whom correspondence should be addressed at: Laboratory ofMetabolism, Center for Cancer Research, National Cancer Institute,National Institutes of Health, 9000 Rockville Pike, Building37, Room 3118, Bethesda, MD 20892. E-mail: kyunglee{at}pop.nci.nih.govor kyunglee{at}mail.nih.gov.
Communicated by Alexander N. Glazer, University of CaliforniaSystem, Oakland, CA, January 28, 2004
Abbreviation: Clb, cyclin B.
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