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Recombination-dependent DNA replication stimulated by double-strand breaks in bacteriophage T4.

Publication ,  Journal Article
Kreuzer, KN; Saunders, M; Weislo, LJ; Kreuzer, HW
Published in: J Bacteriol
December 1995

We analyzed the mechanism of recombination-dependent DNA replication in bacteriophage T4-infected Escherichia coli using plasmids that have sequence homology to the infecting phage chromosome. Consistent with prior studies, a pBR322 plasmid, initially resident in the infected host cell, does not replicate following infection by T4. However, the resident plasmid can be induced to replicate when an integrated copy of pBR322 vector is present in the phage chromosome. As expected for recombination-dependent DNA replication, the induced replication of pBR322 required the phage-encoded UvsY protein. Therefore, recombination-dependent plasmid replication requires homology between the plasmid and phage genomes but does not depend on the presence of any particular T4 DNA sequence on the test plasmid. We next asked whether T4 recombination-dependent DNA replication can be triggered by a double-strand break (dsb). For these experiments, we generated a novel phage strain that cleaves its own genome within the nonessential frd gene by means of the I-TevI endonuclease (encoded within the intron of the wild-type td gene). The dsb within the phage chromosome substantially increased the replication of plasmids that carry T4 inserts homologous to the region of the dsb (the plasmids are not themselves cleaved by the endonuclease). The dsb stimulated replication when the plasmid was homologous to either or both sides of the break but did not stimulate the replication of plasmids with homology to distant regions of the phage chromosome. As expected for recombination-dependent replication, plasmid replication triggered by dsbs was dependent on T4-encoded recombination proteins. These results confirm two important predictions of the model for T4-encoded recombination-dependent DNA replication proposed by Gisela Mosig (p. 120-130, in C. K. Mathews, E. M. Kutter, G. Mosig, and P. B. Berget (ed.), Bacteriophage T4, 1983). In addition, replication stimulated by dsbs provides a site-specific version of the process, which should be very useful for mechanistic studies.

Duke Scholars

Published In

J Bacteriol

DOI

ISSN

0021-9193

Publication Date

December 1995

Volume

177

Issue

23

Start / End Page

6844 / 6853

Location

United States

Related Subject Headings

  • Virus Integration
  • Viral Proteins
  • Sequence Homology, Nucleic Acid
  • Recombination, Genetic
  • Plasmids
  • Mutagenesis
  • Molecular Sequence Data
  • Models, Genetic
  • Microbiology
  • Membrane Proteins
 

Citation

APA
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ICMJE
MLA
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Kreuzer, K. N., Saunders, M., Weislo, L. J., & Kreuzer, H. W. (1995). Recombination-dependent DNA replication stimulated by double-strand breaks in bacteriophage T4. J Bacteriol, 177(23), 6844–6853. https://doi.org/10.1128/jb.177.23.6844-6853.1995
Kreuzer, K. N., M. Saunders, L. J. Weislo, and H. W. Kreuzer. “Recombination-dependent DNA replication stimulated by double-strand breaks in bacteriophage T4.J Bacteriol 177, no. 23 (December 1995): 6844–53. https://doi.org/10.1128/jb.177.23.6844-6853.1995.
Kreuzer KN, Saunders M, Weislo LJ, Kreuzer HW. Recombination-dependent DNA replication stimulated by double-strand breaks in bacteriophage T4. J Bacteriol. 1995 Dec;177(23):6844–53.
Kreuzer, K. N., et al. “Recombination-dependent DNA replication stimulated by double-strand breaks in bacteriophage T4.J Bacteriol, vol. 177, no. 23, Dec. 1995, pp. 6844–53. Pubmed, doi:10.1128/jb.177.23.6844-6853.1995.
Kreuzer KN, Saunders M, Weislo LJ, Kreuzer HW. Recombination-dependent DNA replication stimulated by double-strand breaks in bacteriophage T4. J Bacteriol. 1995 Dec;177(23):6844–6853.

Published In

J Bacteriol

DOI

ISSN

0021-9193

Publication Date

December 1995

Volume

177

Issue

23

Start / End Page

6844 / 6853

Location

United States

Related Subject Headings

  • Virus Integration
  • Viral Proteins
  • Sequence Homology, Nucleic Acid
  • Recombination, Genetic
  • Plasmids
  • Mutagenesis
  • Molecular Sequence Data
  • Models, Genetic
  • Microbiology
  • Membrane Proteins