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Replication of bacteriophage M13: specificity of the Escherichia coli dnaB function for replication of

W L Olsen, W L Staudenbauer, P H Hofschneider

    Proceedings of the National Academy of Sciences of the United States of America
    |September 1, 1972
    PubMed

    Insights

    The dnaB protein is essential for M13 phage DNA replication in E. coli. Without functional dnaB, the phage cannot replicate its double-stranded replicative form (RF) DNA, halting infection.

    Area of Science:

    • Molecular Biology
    • Virology
    • Bacteriology

    Background:

    • Filamentous phage M13 infection in E. coli.
    • Temperature-sensitive E. coli mutant HfrH 165/70 (dnaB).
    • Abortive infections at restrictive temperatures.

    Purpose of the Study:

    • Investigate the role of the dnaB function in M13 phage DNA replication.
    • Determine the specific step in replication dependent on dnaB.
    • Differentiate between single-stranded and double-stranded DNA synthesis requirements.

    Main Methods:

    • Infection of E. coli dnaB mutant with M13 phage at restrictive temperature (41°C).
    • Monitoring phage DNA conversion to replicative form (RF).
    • Observing RF attachment to the cell membrane and subsequent replication.

    Main Results:

    • M13 phage DNA enters the cell and converts to RF at 41°C.
    • Parental RF DNA attaches to the cell membrane.
    • Replication of the double-stranded RF DNA is blocked.
    • Single-stranded DNA synthesis proceeds, but semiconservative RF replication is inhibited.

    Conclusions:

    • The dnaB protein is specifically required for semiconservative replication of the double-stranded M13 RF DNA.
    • This requirement is distinct from the process of single-stranded DNA synthesis.
    • dnaB function is critical for the M13 life cycle in E. coli.

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