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Chromatin and nucleosome structure

R Mandel, G D Fasman

    Nucleic Acids Research
    |August 1, 1976
    PubMed
    Summary
    This summary is machine-generated.

    Chromatin nucleosomes show increased thermal stability with size. Interactions between neighboring nucleosomes in larger structures influence their unfolding and DNA denaturation.

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    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Structural Biology

    Background:

    • Chromatin structure, composed of nucleosomes, is fundamental to DNA organization and regulation.
    • Understanding nucleosome oligomer stability is crucial for elucidating higher-order chromatin structures.

    Purpose of the Study:

    • To investigate the thermal stability and unfolding patterns of isolated chromatin nucleosome oligomers.
    • To determine how nucleosome assembly and size influence DNA denaturation.

    Main Methods:

    • Isolation of mononucleosomes through pentanucleosomes from calf thymus nuclei using staphylococcal nuclease digestion.
    • Circular dichroism spectroscopy to analyze thermal denaturation profiles (Tm) and ellipticity changes (delta[theta]280) as a function of temperature.

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    Main Results:

    • All nucleosome oligomers exhibited similar peak ellipticity, indicating conserved core structure.
    • Melting temperatures (Tm) progressively increased with oligomer size, from 73.5°C (monomer) to 79°C (pentamer).
    • Melting profiles revealed shoulders at 60°C and 93°C, increasing with oligomer size, suggesting complex unfolding beyond simple DNA denaturation.

    Conclusions:

    • Observed thermal denaturation patterns suggest an initial unfolding of asymmetric nucleosome assemblies (polynucleosome superhelix) preceding DNA denaturation.
    • The size-dependent unfolding and increased delta[theta]280 magnitude imply inter-nucleosome interactions in higher-order structures.
    • These findings highlight the role of oligomer size and inter-nucleosome interactions in chromatin stability and DNA accessibility.