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Related Experiment Videos

Human cyclin A is required for mitosis until mid prophase.

N Furuno1, N den Elzen, J Pines

  • 1Wellcome/Cancer Research Campaign Institute and Department of Zoology, University of Cambridge, Cambridge CB2 1QR, United Kingdom.

The Journal of Cell Biology
|October 20, 1999
PubMed
Summary

Cyclin A/cyclin-dependent kinase 2 (CDK2) drives cells into mitosis, but inhibiting it prevents entry. This suggests cyclin A/CDK2 is crucial for cell division progression and may be targeted by the prophase checkpoint.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Mitosis regulation is critical for cell cycle control.
  • Cyclin-dependent kinases (CDKs) are key regulators of cell division.
  • The role of specific cyclin/CDK complexes, like cyclin A/CDK2, in mitosis requires further elucidation.

Purpose of the Study:

  • To investigate the precise role of cyclin A/cyclin-dependent kinase 2 (CDK2) in regulating entry into and progression through mitosis.
  • To determine if cyclin A/CDK2 activity is a rate-limiting step for mitotic entry.
  • To explore the potential involvement of cyclin A/CDK2 as a target of the prophase checkpoint.

Main Methods:

  • Microinjection of purified, active cyclin A/CDK2 into synchronized cells.
  • Time-lapse video microscopy to observe cell division dynamics.

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  • Microinjection of the p21(Cip1/Waf1/Sdi1) amino terminus (p21N) to inhibit cyclin A/CDK2 activity.

    • Main Results:

    • Microinjection of cyclin A/CDK2 induced premature mitosis in G2 cells, often resulting in abnormal cell division.
    • Microinjection of cyclin A/CDK2 into S phase cells did not affect subsequent G2 or mitotic progression.
    • Inhibition of cyclin A/CDK2 by p21N blocked mitotic entry in S and G2 phase cells and caused prophase cells to revert to interphase.

    Conclusions:

    • Cyclin A/CDK2 is a rate-limiting factor essential for initiating mitosis and for progression until late prophase.
    • Cyclin A/CDK2 activity is required for cells to overcome the prophase checkpoint.
    • These findings identify cyclin A/CDK2 as a critical regulator and potential target of the prophase checkpoint mechanism.