This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kreuzer, K. N. Articles by Kreuzer, H. W. Search for Related Content PubMed PubMed Citation Articles by Kreuzer, K. N. Articles by Kreuzer, H. W.

 Previous Article  |  Next Article 

J. Bacteriol., 12 1995, 6844-6853, Vol 177, No. 23
Copyright © 1995, American Society for Microbiology

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

KN Kreuzer, M Saunders, LJ Weislo and HW Kreuzer
Department of Microbiology, Duke University Medical Center, Durham, North Carolina 27710, USA.

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.

This article has been cited by other articles:

• Long, D. T., Kreuzer, K. N. (2008). Regression supports two mechanisms of fork processing in phage T4. Proc. Natl. Acad. Sci. USA 105: 6852-6857 [Abstract] [Full Text]  
• Dudas, K. C., Kreuzer, K. N. (2005). Bacteriophage T4 Helicase Loader Protein gp59 Functions as Gatekeeper in Origin-dependent Replication in Vivo. J. Biol. Chem. 280: 21561-21569 [Abstract] [Full Text]  
• Maisnier-Patin, S., Nordstrom, K., Dasgupta, S. (2001). RecA-Mediated Rescue of Escherichia coli Strains with Replication Forks Arrested at the Terminus. J. Bacteriol. 183: 6065-6073 [Abstract] [Full Text]  
• George, J. W., Stohr, B. A., Tomso, D. J., Kreuzer, K. N. (2001). The tight linkage between DNA replication and double-strand break repair in bacteriophage T4. Proc. Natl. Acad. Sci. USA 98: 8290-8297 [Abstract] [Full Text]  
• Mosig, G., Gewin, J., Luder, A., Colowick, N., Vo, D. (2001). Two recombination-dependent DNA replication pathways of bacteriophage T4, and their roles in mutagenesis and horizontal gene transfer. Proc. Natl. Acad. Sci. USA 98: 8306-8311 [Abstract] [Full Text]  
• Kuzminov, A. (2001). DNA replication meets genetic exchange: Chromosomal damage and its repair by homologous recombination. Proc. Natl. Acad. Sci. USA 98: 8461-8468 [Abstract] [Full Text]  
• Stohr, B. A., Kreuzer, K. N. (2001). Repair of Topoisomerase-Mediated DNA Damage in Bacteriophage T4. Genetics 158: 19-28 [Abstract] [Full Text]  
• Doan, P. L., Belanger, K. G., Kreuzer, K. N. (2001). Two Types of Recombination Hotspots in Bacteriophage T4: One Requires DNA Damage and a Replication Origin and the Other Does Not. Genetics 157: 1077-1087 [Abstract] [Full Text]  
• Valencia-Morales, E., Romero, D. (2000). Recombination Enhancement by Replication (RER) in Rhizobium etli. Genetics 154: 971-983 [Abstract] [Full Text]  
• Parker, M. M., Belisle, M., Belfort, M. (1999). Intron Homing With Limited Exon Homology: Illegitimate Double-Strand-Break Repair in Intron Acquisition by Phage T4. Genetics 153: 1513-1523 [Abstract] [Full Text]  
• Paques, F., Haber, J. E. (1999). Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 63: 349-404 [Abstract] [Full Text]  
• Connelly, J. C., Kirkham, L. A., Leach, D. R. F. (1998). The SbcCD nuclease of Escherichia coli is a structural maintenance of chromosomes (SMC) family protein that cleaves hairpin DNA. Proc. Natl. Acad. Sci. USA 95: 7969-7974 [Abstract] [Full Text]  
• Gary, T. P., Colowick, N. E., Mosig, G. (1998). A Species Barrier Between Bacteriophages T2 and T4: Exclusion, Join-Copy and Join-Cut-Copy Recombination and Mutagenesis in the dCTPase Genes. Genetics 148: 1461-1473 [Abstract] [Full Text]  
• Deng, H., Dewhurst, S. (1998). Functional Identification and Analysis of cis-Acting Sequences Which Mediate Genome Cleavage and Packaging in Human Herpesvirus 6. J. Virol. 72: 320-329 [Abstract] [Full Text]  
• Mueller, J E, Clyman, J, Huang, Y J, Parker, M M, Belfort, M (1996). Intron mobility in phage T4 occurs in the context of recombination-dependent DNA replication by way of multiple pathways.. Genes Dev. 10: 351-364 [Abstract]