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Testing cyclin specificity in the exit from mitosis.

M D Jacobson1, S Gray, M Yuste-Rojas

  • 1The Rockefeller University, New York, NY 10021, USA.

Molecular and Cellular Biology
|June 10, 2000
PubMed
Summary
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Cyclin inactivation is crucial for cell division. Overexpressing a specific cyclin (Clb5p) without its degradation tag causes inviability, while another (Clb2p) causes reversible mitotic arrest, suggesting Clb2p blocks mitosis.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Cyclical inactivation of B-type cyclins is essential for the cell cycle, regulating DNA replication and mitosis.
  • The degradation of Clb5p, mediated by its destruction box, is upregulated during mitosis.

Purpose of the Study:

  • To investigate the role of B-type cyclins, specifically Clb5p and Clb2p, in cell cycle progression.
  • To determine the consequences of constitutive overexpression of cyclins lacking their destruction boxes on cell viability and DNA replication.

Main Methods:

  • Constitutive overexpression of Clb5p and Clb2p lacking their destruction boxes (Deltadb) in yeast cells.
  • Analysis of cell viability, DNA content, and nuclear localization of cyclins.
  • Comparison of phenotypes resulting from CLB5-Deltadb and CLB2-Deltadb overexpression.

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Main Results:

  • Overexpression of CLB5-Deltadb led to inviable cells with variable DNA content, linked to persistent nuclear Clb5p.
  • CLB2-Deltadb overexpression caused a reversible mitotic arrest with uniformly replicated DNA.
  • Simultaneous overexpression resulted in a more reversible mitotic arrest.

Conclusions:

  • Clb2p, not Clb5p, appears to be the primary cyclin that blocks mitotic completion.
  • Persistent high nuclear Clb5p-associated kinase activity may inhibit origin loading, preventing subsequent DNA replication and causing lethality.