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Functional empty capsid precursors produced by lambda mutant defective for late lambda DNA replication.

P Dawson, B Hohn, T Hohn

    Journal of Virology
    |February 1, 1976
    PubMed
    Summary
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    Lambda phage morphogenesis is studied in a mutant with blocked DNA synthesis. Empty head-like structures are identified as normal preheads, awaiting late DNA for completion.

    Area of Science:

    • Bacteriophage biology
    • Molecular genetics
    • Virology

    Background:

    • Lambda phage morphogenesis involves DNA packaging into prehead structures.
    • A specific mutant blocks late phage DNA synthesis, leading to accumulation of early replication products.
    • Previous studies showed late proteins assemble into empty headlike structures in this mutant.

    Purpose of the Study:

    • To characterize the empty headlike structures accumulating in a lambda phage mutant with blocked late DNA synthesis.
    • To determine if these structures are functional precursors to mature phage heads.
    • To investigate the complementation activities present in extracts of the mutant.

    Main Methods:

    • Analysis of lambda phage morphogenesis in a mutant system.
    • Biochemical assays for prehead, tail, and DNA terminigenerating protein activities.

    Related Experiment Videos

  • Sucrose gradient centrifugation of crude extracts.
  • Protein analysis using coelectrophoresis.
  • Electron microscopy of structural components.

    • Main Results:

    • Crude extracts of the mutant contain complementation activities for preheads, tails, and DNA terminigenerating proteins.
    • Sucrose gradients revealed fractions with prehead activity correlating with late protein concentration.
    • Proteins in these fractions matched known prehead capsid proteins.
    • Electron microscopy confirmed the presence of prehead-like structures.

    Conclusions:

    • The empty structures accumulating in the mutant are identified as normal lambda phage preheads.
    • These preheads are capable of assembly but lack the necessary late DNA for filling.
    • The findings provide insight into the regulation of DNA packaging during phage morphogenesis.