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Tests of spool models for DNA packaging in phage lambda.

J Widom, R L Baldwin

    Journal of Molecular Biology
    |December 25, 1983
    PubMed
    Summary
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    This study investigated two models for packaging DNA in phage lambda. Experiments suggest neither the curved-spool nor the straight spool model accurately describes DNA packaging, indicating variability between phages.

    Area of Science:

    • Molecular Biology
    • Virology
    • Biophysics

    Background:

    • Phage lambda DNA packaging models propose DNA is wrapped in cylindrical layers within a spherical capsid.
    • Two models, the curved-spool and straight spool, differ in how DNA segments contact the phage capsid.

    Purpose of the Study:

    • To experimentally test the validity of the curved-spool and straight spool models for phage lambda DNA packaging.
    • To identify specific DNA regions that interact with the phage capsid proteins.

    Main Methods:

    • Chemical crosslinking using ultraviolet irradiation of 5-bromodeoxyuridine-substituted phage lambda.
    • Electron microscopy of lysed phage to visualize DNA-capsid contacts.
    • Restriction endonuclease digestion (EcoRI, HindIII) of crosslinked DNA, followed by density centrifugation and gel electrophoresis to identify crosslinked fragments.

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    Main Results:

    • DNA-capsid contacts were found nearly randomly across the entire phage lambda genome map.
    • Restriction fragments from all genomic regions were identified as being crosslinked to capsid proteins.
    • These findings challenge the proposed specific contact points of both the curved-spool and straight spool models.

    Conclusions:

    • The experimental data largely rule out the curved-spool model and make the straight spool model unlikely, assuming uniform DNA packaging.
    • The results suggest that phage lambda DNA packaging may not be uniform and could vary between individual phage particles.
    • Findings align with other recent research on phage lambda DNA packaging mechanisms.