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Journal of Bacteriology, November 2005, p. 7362-7373, Vol. 187, No. 21
0021-9193/05/$08.00+0     doi:10.1128/JB.187.21.7362-7373.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Type III Restriction Is Alleviated by Bacteriophage (RecE) Homologous Recombination Function but Enhanced by Bacterial (RecBCD) Function

Naofumi Handa1 and Ichizo Kobayashi1,2,3*

Laboratory of Social Genome Sciences, Department of Medical Genome Sciences, Graduate School of Frontier Science,1 Graduate Program in Biophysics and Biochemistry, Graduate School of Science,2 Institute of Medical Science, University of Tokyo, Shirokanedai, Tokyo 108-8639, Japan3

Received 1 April 2005/ Accepted 18 August 2005

Previous works have demonstrated that DNA breaks generated by restriction enzymes stimulate, and are repaired by, homologous recombination with an intact, homologous DNA region through the function of lambdoid bacteriophages lambda and Rac. In the present work, we examined the effect of bacteriophage functions, expressed in bacterial cells, on restriction of an infecting tester phage in a simple plaque formation assay. The efficiency of plaque formation on an Escherichia coli host carrying EcoRI, a type II restriction system, is not increased by the presence of Rac prophage—presumably because, under the single-infection conditions of the plaque assay, a broken phage DNA cannot find a homologue with which to recombine. To our surprise, however, we found that the efficiency of plaque formation in the presence of a type III restriction system, EcoP1 or EcoP15, is increased by the bacteriophage-mediated homologous recombination functions recE and recT of Rac prophage. This type III restriction alleviation does not depend on lar on Rac, unlike type I restriction alleviation. On the other hand, bacterial RecBCD-homologous recombination function enhances type III restriction. These results led us to hypothesize that the action of type III restriction enzymes takes place on replicated or replicating DNA in vivo and leaves daughter DNAs with breaks at nonallelic sites, that bacteriophage-mediated homologous recombination reconstitutes an intact DNA from them, and that RecBCD exonuclease blocks this repair by degradation from the restriction breaks.

* Corresponding author. Mailing address: Laboratory of Social Genome Sciences, Department of Medical Genome Sciences, Graduate School of Frontier Science and Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Phone: (81) 3-5449-5326. Fax: (81) 3-5449-5422. E-mail: ikobaya{at}ims.u-tokyo.ac.jp.

Journal of Bacteriology, November 2005, p. 7362-7373, Vol. 187, No. 21
0021-9193/05/$08.00+0     doi:10.1128/JB.187.21.7362-7373.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Handa, N., Ichige, A., Kobayashi, I. (2009). Contribution of RecFOR machinery of homologous recombination to cell survival after loss of a restriction-modification gene complex. Microbiology 155: 2320-2332 [Abstract] [Full Text]  
  • Ishikawa, K., Handa, N., Kobayashi, I. (2009). Cleavage of a model DNA replication fork by a Type I restriction endonuclease. Nucleic Acids Res 37: 3531-3544 [Abstract] [Full Text]  
  • Yahara, K., Horie, R., Kobayashi, I., Sasaki, A. (2007). Evolution of DNA Double-Strand Break Repair by Gene Conversion: Coevolution Between a Phage and a Restriction-Modification System. Genetics 176: 513-526 [Abstract] [Full Text]  
  • Ivancic-Bace, I., Vlasic, I., Cogelja-Cajo, G., Brcic-Kostic, K., Salaj-Smic, E. (2006). Roles of PriA Protein and Double-Strand DNA Break Repair Functions in UV-Induced Restriction Alleviation in Escherichia coli. Genetics 174: 2137-2149 [Abstract] [Full Text]  


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