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Related Experiment Videos

Conformational constraints in nuclear DNA.

P R Cook, I A Brazell

    Journal of Cell Science
    |November 1, 1976
    PubMed
    Summary
    This summary is machine-generated.

    Researchers studied DNA superstructure in cell nuclei, revealing that nucleoids from most growing cells exhibit supercoiled DNA properties. Gamma-ray irradiation disrupted these DNA structures, while adult hen erythrocytes showed distinct DNA behavior.

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    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Biochemistry

    Background:

    • Cell nuclei contain complex DNA structures.
    • Understanding DNA superstructure is crucial for cellular processes.
    • Nucleoids, protein-depleted nuclear structures, offer a model for studying DNA organization.

    Purpose of the Study:

    • To investigate DNA superstructure in various higher eukaryotic cell nuclei.
    • To analyze the sedimentation properties of nucleoids in the presence of ethidium.
    • To determine the impact of DNA damage on nucleoid structure.

    Main Methods:

    • Gentle cell lysis to isolate nucleoids.
    • Sucrose gradient sedimentation analysis.
    • Ethidium bromide intercalation to assess DNA supercoiling.

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  • Gamma-ray irradiation to induce DNA strand breaks.

    • Main Results:

    • Nucleoids from growing mammalian, avian, amphibian, and insect cells displayed sedimentation patterns characteristic of supercoiled DNA.
    • Gamma-ray irradiation, causing single-strand breaks, abolished these characteristic sedimentation changes.
    • Nucleoids from adult hen erythrocytes showed no variation in sedimentation rate, differing from other cell types.

    Conclusions:

    • Growing cells' DNA exhibits supercoiling, influencing nucleoid sedimentation.
    • DNA integrity, specifically supercoiling, is essential for the observed sedimentation properties.
    • Adult hen erythrocyte nucleoids possess a unique DNA superstructure distinct from other studied cell types.