Repair of Double-Strand Breaks in Bacteriophage T4 by a Mechanism That Involves Extensive DNA Replication

INVESTIGATIONS

Repair of Double-Strand Breaks in Bacteriophage T4 by a Mechanism That Involves Extensive DNA Replication

J. W. George and K. N. Kreuzer
Department of Microbiology, Duke University Medical Center, Durham, North Carolina 27710

We investigated double-strand break (dsb) repair in bacteriophage T4 using a physical assay that involves a plasmid substrate with two inverted DNA segments. A dsb introduced into one repeat during a T4 infection induces efficient dsb repair using the second repeat as a template. This reaction is characterized by the following interesting features. First, the dsb induces a repair reaction that is directly coupled to extensive plasmid replication; the repaired/replicated product is in the form of long plasmid concatemers. Second, repair of the dsb site is frequently associated with exchange of flanking DNA. Third, the repair reaction is absolutely dependent on the products of genes uvsX, uvsY, 32, 46, and 59, which are also required for phage genomic recombination-dependent DNA replication. Fourth, the coupled repair/replication reaction is only partly dependent on endonuclease VII (gp49), suggesting that either another Holliday-junction-cleaving activity or an alternate resolution pathway is active during T4 infections. Because this repair reaction is directly coupled to extensive replication, it cannot be explained by the SZOSTAK et al. model. We present and discuss a model for the coupled repair/replication reaction, called the extensive chromosome replication model for dsb repair.

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				B. A. Stohr and K. N. Kreuzer Coordination of DNA Ends During Double-Strand-Break Repair in Bacteriophage T4 Genetics, November 1, 2002; 162(3): 1019 - 1030. [Abstract] [Full Text] [PDF]

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				J. W. George, B. A. Stohr, D. J. Tomso, and K. N. Kreuzer The tight linkage between DNA replication and double-strand break repair in bacteriophage T4 PNAS, July 17, 2001; 98(15): 8290 - 8297. [Abstract] [Full Text] [PDF]

				J. S. Bleuit, H. Xu, Y. Ma, T. Wang, J. Liu, and S. W. Morrical Mediator proteins orchestrate enzyme-ssDNA assembly during T4 recombination-dependent DNA replication and repair PNAS, July 17, 2001; 98(15): 8298 - 8305. [Abstract] [Full Text] [PDF]

				G. Mosig, J. Gewin, A. Luder, N. Colowick, and D. Vo Two recombination-dependent DNA replication pathways of bacteriophage T4, and their roles in mutagenesis and horizontal gene transfer PNAS, July 17, 2001; 98(15): 8306 - 8311. [Abstract] [Full Text] [PDF]

				C. E. Jones, T. C. Mueser, K. C. Dudas, K. N. Kreuzer, and N. G. Nossal Bacteriophage T4 gene 41 helicase and gene 59 helicase-loading protein: A versatile couple with roles in replication and recombination PNAS, July 17, 2001; 98(15): 8312 - 8318. [Abstract] [Full Text] [PDF]

				B. A. Stohr and K. N. Kreuzer Repair of Topoisomerase-Mediated DNA Damage in Bacteriophage T4 Genetics, May 1, 2001; 158(1): 19 - 28. [Abstract] [Full Text]

				M. Yu and W. Masker T7 Single Strand DNA Binding Protein but Not T7 Helicase Is Required for DNA Double Strand Break Repair J. Bacteriol., March 15, 2001; 183(6): 1862 - 1869. [Abstract] [Full Text]

				L. Signon, A. Malkova, M. L. Naylor, H. Klein, and J. E. Haber Genetic Requirements for RAD51- and RAD54-Independent Break-Induced Replication Repair of a Chromosomal Double-Strand Break Mol. Cell. Biol., March 15, 2001; 21(6): 2048 - 2056. [Abstract] [Full Text]

				J. M. Pluth, L. M. Fried, and C. U. Kirchgessner Severe Combined Immunodeficient Cells Expressing Mutant hRAD54 Exhibit a Marked DNA Double-Strand Break Repair and Error-prone Chromosome Repair Defect Cancer Res., March 1, 2001; 61(6): 2649 - 2655. [Abstract] [Full Text]

				P. L. Doan, K. G. Belanger, and K. N. Kreuzer Two Types of Recombination Hotspots in Bacteriophage T4: One Requires DNA Damage and a Replication Origin and the Other Does Not Genetics, March 1, 2001; 157(3): 1077 - 1087. [Abstract] [Full Text]

				D. J. Tomso and K. N. Kreuzer Double-Strand Break Repair in Tandem Repeats During Bacteriophage T4 Infection Genetics, August 1, 2000; 155(4): 1493 - 1504. [Abstract] [Full Text]

				Y.-T. Lai and W. Masker Visualization of Repair of Double-Strand Breaks in the Bacteriophage T7 Genome without Normal DNA Replication J. Bacteriol., January 15, 2000; 182(2): 327 - 336. [Abstract] [Full Text]

				G. Hong and K. N. Kreuzer An Antitumor Drug-Induced Topoisomerase Cleavage Complex Blocks a Bacteriophage T4 Replication Fork In Vivo Mol. Cell. Biol., January 15, 2000; 20(2): 594 - 603. [Abstract] [Full Text]

				Y.-J. Huang, M. M. Parker, and M. Belfort Role of Exonucleolytic Degradation in Group I Intron Homing in Phage T4 Genetics, December 1, 1999; 153(4): 1501 - 1512. [Abstract] [Full Text]

				M. M. Parker, M. Belisle, and M. Belfort Intron Homing With Limited Exon Homology: Illegitimate Double-Strand-Break Repair in Intron Acquisition by Phage T4 Genetics, December 1, 1999; 153(4): 1513 - 1523. [Abstract] [Full Text]

				M. P. Colaiácovo, F. Pâques, and J. E. Haber Removal of One Nonhomologous DNA End During Gene Conversion by a RAD1- and MSH2-Independent Pathway Genetics, April 1, 1999; 151(4): 1409 - 1423. [Abstract] [Full Text]

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				T. P. Gary, N. E. Colowick, and G. Mosig A Species Barrier Between Bacteriophages T2 and T4: Exclusion, Join-Copy and Join-Cut-Copy Recombination and Mutagenesis in the dCTPase Genes Genetics, April 1, 1998; 148(4): 1461 - 1473. [Abstract] [Full Text] [PDF]

				J E Mueller, D Smith, and M Belfort Exon coconversion biases accompanying intron homing: battle of the nucleases. Genes & Dev., September 1, 1996; 10(17): 2158 - 2166. [Abstract] [PDF]

				C. E. Jones, T. C. Mueser, and N. G. Nossal Interaction of the Bacteriophage T4 Gene 59 Helicase Loading Protein and Gene 41 Helicase with Each Other and with Fork, Flap, and Cruciform DNA J. Biol. Chem., August 25, 2000; 275(35): 27145 - 27154. [Abstract] [Full Text] [PDF]