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Involvement of Escherichia coli K-12 DNA polymerase I in the growth of bacteriophage Mu

Journal of Virology
|October 1, 1983
PubMed

Insights

Mutations in the polA gene, which codes for DNA polymerase I (PolI), significantly impair bacteriophage Mu development in Escherichia coli. PolI is crucial for Mu lysogenization, lytic growth, and transposition.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Bacteriophage Mu is a versatile genetic element used extensively in molecular biology.
  • DNA polymerase I (PolI) is a key enzyme in DNA replication and repair in Escherichia coli.

Purpose of the Study:

  • To investigate the role of DNA polymerase I (PolI) in the development of bacteriophage Mu in Escherichia coli.
  • To determine the impact of polA gene mutations on Mu lysogenization, lytic growth, and transposition.

Main Methods:

  • Comparative analysis of bacteriophage Mu development in polA mutant and wild-type Escherichia coli strains.
  • Lysogenization frequency assays.
  • One-step growth experiments.
  • Mu-mediated transposition assays.
  • Pulse labeling and DNA-DNA hybridization for Mu DNA synthesis analysis.

Main Results:

  • polA mutants showed a 3-to-8-fold lower frequency of Mu lysogenization compared to wild-type strains.
  • Phage Mu exhibited a longer latent period (50-100% increase) and reduced burst size (20-40% decrease) in polA mutants.
  • A 10-fold reduction in Mu-mediated transposition of chromosomal genes to an F plasmid was observed in polA strains.
  • Mu DNA replication initiated similarly but proceeded at a slower rate in polA cells.

Conclusions:

  • DNA polymerase I (PolI) plays a significant role in the replication and integration of bacteriophage Mu.
  • Phage Mu development is impaired but not completely abolished in the absence of normal PolI activity, suggesting partial compensation by residual activity or other enzymes.

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