Requirement of Cdk2-cyclin E activity for repeated centrosome reproduction in Xenopus egg extracts
Hinchcliffe, Edward H. ; Li, Chuan ; Thompson, Elizabeth A. ; Maller, James L. ; Sluder, Greenfield
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UMass Chan Affiliations
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Keywords
Aphidicolin
Blastomeres
*CDC2-CDC28 Kinases
*Cell Cycle Proteins
Cell Extracts
Centrosome
Cyclin E
Cyclin-Dependent Kinase 2
Cyclin-Dependent Kinase Inhibitor p27
Cyclin-Dependent Kinases
DNA
Enzyme Inhibitors
Microscopy, Confocal
Microscopy, Fluorescence
Microscopy, Video
Microtubule-Associated Proteins
Ovum
Protein Kinases
Protein-Serine-Threonine Kinases
Proteins
Recombinant Proteins
*S Phase
*Tumor Suppressor Proteins
Xenopus
Xenopus Proteins
Cell Biology
Life Sciences
Medicine and Health Sciences
Subject Area
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Abstract
The abnormally high number of centrosomes found in many human tumor cells can lead directly to aneuploidy and genomic instability through the formation of multipolar mitotic spindles. To facilitate investigation of the mechanisms that control centrosome reproduction, a frog egg extract arrested in S phase of the cell cycle that supported repeated assembly of daughter centrosomes was developed. Multiple rounds of centrosome reproduction were blocked by selective inactivation of cyclin-dependent kinase 2-cyclin E (Cdk2-E) and were restored by addition of purified Cdk2-E. Confocal immunomicroscopy revealed that cyclin E was localized at the centrosome. These results demonstrate that Cdk2-E activity is required for centrosome duplication during S phase and suggest a mechanism that could coordinate centrosome reproduction with cycles of DNA synthesis and mitosis.
Source
Science. 1999 Feb 5;283(5403):851-4.