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