Phosphorylation of Centrin during the Cell Cycle and Its Role in Centriole Separation Preceding Centrosome Duplication

Ward Lutz, Wilma L. Lingle, Daniel J Mc Cormick, Tammy M. Greenwood, Jeffrey L Salisbury

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Abstract

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 G 2/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.

Original languageEnglish (US)
Pages (from-to)20774-20780
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number23
DOIs
StatePublished - Jun 8 2001

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Centrioles
Centrosome
Phosphorylation
Sulfamethoxazole Drug Combination Trimethoprim
Cell Cycle
Cells
Spindle Poles
Prophase
Spindle Apparatus
Interphase
Staining and Labeling
Labeling
Poles
Telophase
Anaphase
Antibodies
Metaphase
Indirect Fluorescent Antibody Technique
HeLa Cells
Cell Division

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation of Centrin during the Cell Cycle and Its Role in Centriole Separation Preceding Centrosome Duplication. / Lutz, Ward; Lingle, Wilma L.; Mc Cormick, Daniel J; Greenwood, Tammy M.; Salisbury, Jeffrey L.

In: Journal of Biological Chemistry, Vol. 276, No. 23, 08.06.2001, p. 20774-20780.

Research output: Contribution to journalArticle

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AB - 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 G 2/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.

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