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Knotted DNA from bacteriophage capsids.

L F Liu, L Perkocha, R Calendar

    Proceedings of the National Academy of Sciences of the United States of America
    |September 1, 1981
    PubMed
    Summary
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    DNA from tailless bacteriophage P2 capsids forms complex knots when cohesive ends join. This knotted DNA structure can be utilized to assay type II DNA topoisomerases, enzymes crucial for DNA manipulation.

    Area of Science:

    • Molecular Biology
    • Virology
    • Biochemistry

    Background:

    • Bacteriophage P2 DNA isolated from tailless capsids exhibits unusual properties.
    • The phenol extraction procedure yields predominantly circular DNA molecules with joined cohesive ends.

    Purpose of the Study:

    • To investigate the complex structure of DNA from tailless P2 capsids.
    • To determine the topological state of the DNA and its implications.
    • To explore the potential use of this DNA in enzyme assays.

    Main Methods:

    • Electron microscopy and ultracentrifugation to analyze DNA structure.
    • DNA manipulation techniques including thermal dissociation and enzymatic repair (DNA polymerase I).
    • Enzymatic treatments with ligase and DNase I to probe DNA topology.

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    Main Results:

    • The majority of P2 DNA from tailless capsids exists as topologically knotted monomeric rings.
    • These knotted DNA molecules display a more compact structure and higher sedimentation coefficient than regular P2 DNA rings.
    • Linearization and enzymatic treatments confirm the knotted nature and allow for comparison with normal P2 DNA.

    Conclusions:

    • DNA packaging in tailless P2 capsids leads to the formation of complex DNA knots upon end joining.
    • The presence of a tail in intact phage particles likely prevents such knot formation.
    • The unique knotted DNA structure serves as a substrate for assaying type II DNA topoisomerases.