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A protein homologous to human Ku p70-protein is required for reconstitution ofXenopus sperm pronuclei

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Abstract

The monoclonal antibody mAbH6, which recognizes one human Ku-protein (p70), cross-reacted with the counterpart protein fromXenopus eggs. TheXenopus antigen purified with a mAbH6-Sepharose column was a complex of 88 kDa and 72 kDa proteins. The role of Ku protein in nuclear structure formation was studied using a cell-free nuclear assembly extract derived fromXenopus interphase eggs. The protein was distributed on the surface of demembranated sperm chromatin and in the membrane vesicle fraction of the nuclear assembly extract. Addition of mAbH6 to the assembly extracts prevented the completion of reconstitution of pronuclei from demembranated sperm chromatins, although partial decondensation mediated by nucleoplasmin was not impeded. As a result, the sperm chromatin remained in partially swollen structures or formed round but small anomalous nuclei. Nuclear membranes were formed on the nuclei in mAbH6-inhibited extract systems, but DNA synthesis was largely decreased, suggesting the incomplete reconstitution of pronuclei. The incorporation of lamin to the nuclei was inhibited by mAbH6. It is suggested that theXenopus Ku-homologous protein has a role in the formation of the lamin layer after nuclear membrane reconstitution.

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Authors and Affiliations

  1. the Department of Biology, Faculty of Science, Kwansei Gakuin University, 662, Nishinomiya, Hyogo, Japan

    Masahito Higashiura, Yoshihiro Takasuga & Tatsuo Yagura

  2. the Radio Isotope Research Center, Osaka University, 565, Suita, Osaka, Japan

    Jinpei Yamashita

Authors
  1. Masahito Higashiura
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  2. Yoshihiro Takasuga
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  3. Jinpei Yamashita
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  4. Tatsuo Yagura
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Higashiura, M., Takasuga, Y., Yamashita, J. et al. A protein homologous to human Ku p70-protein is required for reconstitution ofXenopus sperm pronuclei. Chromosome Res 1, 27–36 (1993). https://doi.org/10.1007/BF00710604

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  • DOI: https://doi.org/10.1007/BF00710604

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