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Related Experiment Videos

Nucleosome conformation: pH and organic solvent effects.

M Zama, D E Olins, B Prescott

    Nucleic Acids Research
    |October 1, 1978
    PubMed
    Summary

    Monomer nucleosomes undergo conformational changes in response to pH and solvent variations. These alterations affect DNA structure and histone alpha-helix content, impacting nucleosome stability.

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    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Structural Biology

    Background:

    • Nucleosomes are fundamental units of DNA packaging in eukaryotes.
    • Understanding nucleosome conformational dynamics is crucial for gene regulation.

    Purpose of the Study:

    • To investigate the conformational transitions of monomer nucleosomes (nu 1) under varying pH and solvent conditions.
    • To elucidate the structural changes in both DNA and histone components of nucleosomes.

    Main Methods:

    • Circular dichroism spectroscopy to assess DNA and histone secondary structures.
    • Laser Raman spectroscopy to probe nucleosome conformation.
    • Fluorescence spectroscopy of pyrene-labeled histone H3 to monitor structural changes.

    Main Results:

    • pH-dependent transitions observed: DNA ellipticity suppressed (pH 5.5-4.8), irreversible B-DNA character increase and histone alpha-helix loss (pH 4.6-4.2), nucleosome aggregation (pH < 4).
    • Solvent mixtures induced similar conformational changes, including increased B-DNA character and histone alpha-helix reduction.
    • Pyrene excimer fluorescence loss correlated with structural transitions, indicating conformational alterations.

    Conclusions:

    • Monomer nucleosomes exhibit distinct conformational transitions in response to acidic pH and various organic solvents.
    • These transitions involve significant alterations in DNA conformation and histone protein structure.
    • The findings provide insights into the stability and structural plasticity of nucleosomes.

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