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Salt-induced structural changes in nucleosomes

C G Sahasrabuddhe, G F Saunders

    Nucleic Acids Research
    |April 1, 1977
    PubMed
    Summary
    This summary is machine-generated.

    Increasing salt concentration causes nucleosomes to condense and become insoluble, affecting their structure. This insolubility allows for analysis of nucleosomal DNA changes using spectroscopy.

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    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Structural Biology

    Background:

    • Nucleosomes are the basic units of DNA packaging in eukaryotes.
    • Understanding nucleosome structure is crucial for comprehending gene regulation and DNA accessibility.

    Purpose of the Study:

    • To investigate the structural changes of nucleosomes and oligonucleosomes under varying salt concentrations.
    • To determine the salt concentration range for nucleosome insolubility and its implications for structural analysis.

    Main Methods:

    • Preparation of nucleosomes and oligonucleosomes from human placental nuclei using staphylococcal nuclease digestion.
    • Fractionation of nucleosomes and oligonucleosomes by gel filtration chromatography.
    • Analysis of structural changes using Circular Dichroism (C.D.) spectroscopy and turbidity measurements in the presence and absence of MgCl2.

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    Main Results:

    • Nucleosomes and oligonucleosomes exhibit insolubility across a wide range of salt concentrations.
    • Nucleosome insolubility is observed at or above 120 mM (NH4)2SO4 with 10 mM MgCl2, enabling C.D. spectroscopic analysis.
    • Turbidity measurements confirm nucleosome insolubility at or below 120 mM (NH4)2SO4 with 10 mM MgCl2.

    Conclusions:

    • Nucleosome insolubility is linked to salt-induced structural alterations, likely due to particle condensation.
    • Nucleosomes undergo condensation at increasing salt concentrations, followed by relaxation at higher concentrations.