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Related Experiment Videos

Chromosome-bound mitotic factors: release by endonucleases.

R C Adlakha, C G Sahasrabuddhe, D A Wright

    Nucleic Acids Research
    |July 10, 1982
    PubMed
    Summary

    Mammalian mitotic factors, crucial for cell division, are confirmed to be located on metaphase chromosomes. Mild enzymatic digestion partially released these factors, supporting their chromosomal localization.

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    Area of Science:

    • Cell Biology
    • Molecular Biology
    • Developmental Biology

    Background:

    • Mammalian mitotic factors induce germinal vesicle breakdown and chromosome condensation in Xenopus laevis oocytes.
    • Previous research suggested these factors are localized on metaphase chromosomes.

    Purpose of the Study:

    • To provide additional evidence for the localization of mammalian mitotic factors on metaphase chromosomes.
    • To investigate the release of these factors from chromosomes using enzymatic digestion.

    Main Methods:

    • Mitotic HeLa cell chromosomes were purified via sucrose gradients.
    • Chromosomes were digested with micrococcal nuclease or DNase II.
    • Released factors were quantified by injecting oocyte supernatants and assessing oocyte responses.

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    Main Results:

    • Mild enzymatic digestion (3-5% DNA solubilized) released 30-40% of mitotic factors.
    • Re-extraction of the chromosomal pellet with 0.2 M NaCl confirmed factor release.
    • Similar results were observed with G2 cell nuclei.

    Conclusions:

    • A significant proportion of mammalian mitotic factors are indeed bound to metaphase chromosomes.
    • Enzymatic digestion provides a method to partially release and study these chromosome-bound factors.