Requirement of Cdk2-cyclin E activity for repeated centrosome reproduction in Xenopus egg extracts

Science. 1999 Feb 5;283(5403):851-4. doi: 10.1126/science.283.5403.851.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Aphidicolin / pharmacology
  • Blastomeres / chemistry
  • CDC2-CDC28 Kinases*
  • Cell Cycle Proteins*
  • Cell Extracts
  • Centrosome / chemistry
  • Centrosome / metabolism*
  • Cyclin E / analysis
  • Cyclin E / metabolism*
  • Cyclin-Dependent Kinase 2
  • Cyclin-Dependent Kinase Inhibitor p27
  • Cyclin-Dependent Kinases / antagonists & inhibitors
  • Cyclin-Dependent Kinases / metabolism*
  • DNA / biosynthesis
  • Enzyme Inhibitors / pharmacology
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Microscopy, Video
  • Microtubule-Associated Proteins / pharmacology
  • Ovum
  • Protein Kinases / metabolism
  • Protein Serine-Threonine Kinases / antagonists & inhibitors
  • Protein Serine-Threonine Kinases / metabolism*
  • Proteins
  • Recombinant Proteins / pharmacology
  • S Phase*
  • Tumor Suppressor Proteins*
  • Xenopus
  • Xenopus Proteins

Substances

  • Cell Cycle Proteins
  • Cell Extracts
  • Cyclin E
  • Enzyme Inhibitors
  • Microtubule-Associated Proteins
  • Proteins
  • Recombinant Proteins
  • Tumor Suppressor Proteins
  • Xenopus Proteins
  • Xicl protein, Xenopus
  • Cyclin-Dependent Kinase Inhibitor p27
  • Aphidicolin
  • DNA
  • Protein Kinases
  • histone H1 kinase
  • Protein Serine-Threonine Kinases
  • CDC2-CDC28 Kinases
  • Cdk2 protein, Xenopus
  • Cyclin-Dependent Kinase 2
  • Cyclin-Dependent Kinases