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cdc2 phosphorylation is required for its interaction with cyclin.

B Ducommun1, P Brambilla, M A Félix

  • 1Differentiation Program, European Molecular Biology Laboratory, Heidelberg, FRG.

The EMBO Journal
|November 11, 1991
PubMed
Summary
This summary is machine-generated.

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Phosphorylation of the cdc2 protein is crucial for its interaction with cyclins, regulating cell cycle progression. Specific mutations, like Thr161Val, disrupt this process, leading to cell cycle defects and lethality.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Cell cycle progression is tightly regulated by the activation of cyclin-dependent kinases, such as cdc2.
  • Post-translational modifications and cyclin interactions are key regulatory mechanisms for cdc2 protein kinase activity.

Purpose of the Study:

  • To investigate the role of post-translational modifications, specifically phosphorylation at Thr161, in regulating cdc2 binding to cyclins A and B.
  • To elucidate the impact of specific cdc2 mutations on cyclin binding and cellular function.

Main Methods:

  • In vitro translation of human cdc2.
  • Preincubation with Xenopus egg extract to induce phosphorylation.
  • Site-directed mutagenesis of cdc2 (Thr161Val, Thr161Glu, N-terminus, PSTAIR region).

Related Experiment Videos

  • Assessment of cdc2-cyclin binding affinity.
  • Phenotypic analysis in Schizosaccharomyces pombe.

    • Main Results:

    • Preincubation with Xenopus egg extract induced cdc2 phosphorylation, enabling binding to cyclins A and B.
    • Mutation of Thr161 to valine (Thr161Val) abolished cdc2-cyclin binding and caused lethality in S. pombe.
    • Mutations in the N-terminus and PSTAIR region also inhibited cdc2-cyclin binding.
    • Mimicking phosphorylation at Thr161 with glutamic acid (Thr161Glu) resulted in mitotic and cytokinesis defects.

    Conclusions:

    • Phosphorylation of cdc2 at Thr161 is essential for its association with cyclins A and B.
    • A threonine phosphorylation/dephosphorylation cycle regulates cdc2 function during the cell cycle.
    • Specific mutations in cdc2 can lead to severe cell cycle dysregulation and organismal lethality.