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Xenopus laevis actin-depolymerizing factor/cofilin: a phosphorylation-regulated protein essential for development

H Abe1, T Obinata, L S Minamide

  • 1Department of Biology, Chiba University, Japan.

The Journal of Cell Biology
|March 1, 1996
PubMed
Summary
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Xenopus actin-depolymerizing factors (XADs) are crucial for cell division. Proper regulation of XAD activity is essential for cytokinesis during early embryonic development.

Area of Science:

  • Cell Biology
  • Developmental Biology
  • Biochemistry

Background:

  • Two cDNAs encoding Xenopus ADF/cofilins (XADs) were identified from a Xenopus laevis embryonic library.
  • These proteins share high sequence identity with chick ADF and cofilin, known regulators of actin dynamics.

Purpose of the Study:

  • To investigate the function and regulation of XAD proteins during Xenopus laevis embryonic development.
  • To determine the role of XADs in cytokinesis and early embryonic cell division.

Main Methods:

  • Isolation and characterization of XAD cDNAs.
  • Purification of GST-fusion proteins to assess actin-depolymerizing and binding activities.
  • Immunofluorescence localization of XAC in oocytes and embryos.
  • 2D Western blotting to identify phosphorylated forms (pXAC).

Related Experiment Videos

  • Functional assays involving injection of active XAC or inhibitory antibodies into blastomeres.

    • Main Results:

    • XAD proteins exhibit pH-dependent actin-depolymerizing and F-actin-binding activities.
    • XAC is localized in the cortical cytoplasm of oocytes and dynamically redistributed after fertilization, concentrating at the membrane and cleavage furrow.
    • A phosphorylated form (pXAC) is prevalent in oocytes and rapidly dephosphorylated post-fertilization.
    • Modulating XAC activity (constitutively active or inhibitory antibody) specifically blocked injected blastomere cleavage without affecting nuclear division, demonstrating a role in cytokinesis.

    Conclusions:

    • Xenopus ADF/cofilins (XADs) play a critical role in regulating cytokinesis during early embryonic development.
    • The activity of XADs must be tightly regulated, likely through phosphorylation/dephosphorylation, for successful cell cleavage.
    • XADs are essential components of the contractile machinery required for cell division in Xenopus embryos.