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Association of p34cdc2/cyclin B complex with microtubules in starfish oocytes

K Ookata1, S Hisanaga, E Okumura

  • 1Laboratory of Cell and Developmental Biology, Faculty of Biosciences, Tokyo Institute of Technology, Yokohama, Japan.

Journal of Cell Science
|August 1, 1993
PubMed

Insights

Maturation promoting factor (MPF), a complex of p34cdc2 and cyclin B, directly associates with microtubules. This interaction, mediated by microtubule-associated proteins (MAPs), is crucial for microtubule reorganization during cell division.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Cytoskeleton Dynamics

Background:

  • The microtubular cytoskeleton undergoes significant reorganization from interphase to the mitotic spindle during the G2/M transition.
  • Maturation promoting factor (MPF), comprising p34cdc2 and cyclin B, is implicated in this reorganization, but its precise role in controlling microtubule networks is unclear.

Purpose of the Study:

  • To investigate the direct association of the p34cdc2/cyclin B complex with microtubules.
  • To elucidate the mechanism by which p34cdc2 kinase influences microtubule organization.

Main Methods:

  • Immunofluorescence staining of starfish oocytes, asters, and meiotic spindles for anti-cyclin B.
  • Biochemical analysis of starfish oocyte microtubules, including high-salt extraction, suc1-bead adsorption, and immunoprecipitation.
  • In vitro binding assays using purified p34cdc2/cyclin B, porcine brain microtubules, and starfish oocyte MAPs.

Main Results:

  • Direct association of p34cdc2/cyclin B complex with microtubules, centrosomes, and chromosomes was observed.
  • Microtubule-bound p34cdc2 and cyclin B were released by high salt and confirmed as a complex.
  • The p34cdc2/cyclin B complex bound to microtubules in both active (meiotic metaphase) and inactive (immature oocytes) forms.
  • In vitro experiments demonstrated that MAPs mediate the association of p34cdc2/cyclin B with microtubules.

Conclusions:

  • The p34cdc2/cyclin B complex directly binds to microtubules in starfish oocytes.
  • This association is mediated by MAPs and occurs independently of the kinase activity of p34cdc2.
  • These findings provide a molecular basis for MPF's role in regulating microtubule dynamics during cell division.

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