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Structures and dynamics of a supercoiled DNA.

J H Shibata, J Wilcoxon, J M Schurr

    Biochemistry
    |March 13, 1984
    PubMed
    Summary

    Supercoiled DNA exhibits distinct structural conformers with unique torsional rigidities, suggesting non-B helical secondary structures. These findings offer insights into DNA

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    Area of Science:

    • Molecular Biology
    • Biophysics
    • Genetics

    Background:

    • Supercoiled DNA plays a crucial role in various biological processes, including replication and gene regulation.
    • Understanding the structural dynamics of supercoiled DNA is essential for deciphering its functional mechanisms.

    Purpose of the Study:

    • To investigate the secondary and tertiary structures of supercoiled M13mp7 DNA.
    • To identify and characterize different structural conformers and their properties.
    • To explore the implications of these structures for DNA function and gene regulation.

    Main Methods:

    • Time-resolved fluorescence polarization anisotropy to monitor torsional rigidity.
    • Gel electrophoresis to assess tertiary structures and conformer separation.

    Main Results:

    • Identification of seven distinct, long-lived structural conformers of supercoiled M13mp7 DNA.
    • Two prevalent secondary structures were observed, differing from the canonical B-helix.
    • Conformers exhibited distinct torsional rigidities and gel mobilities, influenced by buffer conditions and preparation methods.

    Conclusions:

    • Supercoiled DNA can adopt at least two distinct secondary structures, suggesting a regulatory role beyond simple helix formation.
    • The observed metastability and hysteresis support a model where supercoiling facilitates remote gene control via protein interactions.
    • A model is proposed for gene regulation involving protein-mediated control of secondary structure domain junctions, explaining phenomena like the prmup-1 mutation's effect.

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