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Science 332 (6037): 1557-1561

Copyright © 2011 by the American Association for the Advancement of Science

A Cell Cycle Phosphoproteome of the Yeast Centrosome

Jamie M. Keck,1,* Michele H. Jones,1,* Catherine C. L. Wong,2 Jonathan Binkley,3 Daici Chen,4 Sue L. Jaspersen,5,6 Eric P. Holinger,1 Tao Xu,2 Mario Niepel,7 Michael P. Rout,8 Jackie Vogel,4 Arend Sidow,3 John R. Yates, III,2 Mark Winey1,{dagger}

Abstract: Centrosomes organize the bipolar mitotic spindle, and centrosomal defects cause chromosome instability. Protein phosphorylation modulates centrosome function, and we provide a comprehensive map of phosphorylation on intact yeast centrosomes (18 proteins). Mass spectrometry was used to identify 297 phosphorylation sites on centrosomes from different cell cycle stages. We observed different modes of phosphoregulation via specific protein kinases, phosphorylation site clustering, and conserved phosphorylated residues. Mutating all eight cyclin-dependent kinase (Cdk)–directed sites within the core component, Spc42, resulted in lethality and reduced centrosomal assembly. Alternatively, mutation of one conserved Cdk site within {gamma}-tubulin (Tub4-S360D) caused mitotic delay and aberrant anaphase spindle elongation. Our work establishes the extent and complexity of this prominent posttranslational modification in centrosome biology and provides specific examples of phosphorylation control in centrosome function.

1 Department of Molecular, Cellular, and Developmental Biology, UCB 347, University of Colorado, Boulder, CO 80309, USA.
2 Department of Chemical Physiology, Scripps Research Institute, La Jolla, CA 92037, USA.
3 Departments of Pathology and Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
4 Department of Biology, McGill University, Montreal, Quebec H3G 0B1, Canada.
5 Stowers Institute for Medical Research, Kansas City, MO 64110, USA.
6 Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
7 Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
8 Laboratory of Cellular and Structural Biology, Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.

* These authors contributed equally to this work.

{dagger} To whom correspondence should be addressed. E-mail: mark.winey{at}colorado.edu


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