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Originally published In Press as doi:10.1074/jbc.M101324200 on March 12, 2001

J. Biol. Chem., Vol. 276, Issue 23, 20774-20780, June 8, 2001
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Phosphorylation of Centrin during the Cell Cycle and Its Role in Centriole Separation Preceding Centrosome Duplication*

Ward Lutz, Wilma L. Lingle, Daniel McCormick, Tammy M. Greenwood, and Jeffrey L. SalisburyDagger

From the Department of Biochemistry and Molecular Biology, Tumor Biology Program, Mayo Clinic Foundation, Rochester, Minnesota 55905

Once during each cell cycle, mitotic spindle poles arise by separation of newly duplicated centrosomes. We report here the involvement of phosphorylation of the centrosomal protein centrin in this process. We show that centrin is phosphorylated at serine residue 170 during the G2/M phase of the cell cycle. Indirect immunofluorescence staining of HeLa cells using a phosphocentrin-specific antibody reveals intense labeling of mitotic spindle poles during prophase and metaphase of the cell division cycle, with diminished staining of anaphase and no staining of telophase and interphase centrosomes. Cultured cells undergo a dramatic increase in centrin phosphorylation following the experimental elevation of PKA activity, suggesting that this kinase can phosphorylate centrin in vivo. Surprisingly, elevated PKA activity also resulted intense phosphocentrin antibody labeling of interphase centrosomes and in the concurrent movement of individual centrioles apart from one another. Taken together, these results suggest that centrin phosphorylation signals the separation of centrosomes at prophase and implicates centrin phosphorylation in centriole separation that normally precedes centrosome duplication.

* This work was supported by NCI, National Institutes of Health Grant CA72836 (to J. L. S.), Department of Defense Grant DAMD17-98-1-8122 (to W. L. L.), and the Mayo Clinic Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Tumor Biology Program, Mayo Clinic Foundation, Rochester, MN 55905. Tel.: 507-284-3326; Fax: 507-284-1767; E-mail: salisbury@mayo.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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