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Journal of Bacteriology, June 2009, p. 3785-3793, Vol. 191, No. 12
0021-9193/09/$08.00+0     doi:10.1128/JB.01817-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Functional Characterization of Excision Repair and RecA-Dependent Recombinational DNA Repair in Campylobacter jejuni{triangledown}

Esther J. Gaasbeek,1,2 Fimme J. van der Wal,1,3 Jos P. M. van Putten,2,3 Paulo de Boer,1,{ddagger} Linda van der Graaf-van Bloois,1,3 Albert G. de Boer,1,3 Bart J. Vermaning,1 and Jaap A. Wagenaar1,2,3*

Department of Bacteriology and TSEs, Central Veterinary Institute of Wageningen UR, 8219 PH Lelystad, The Netherlands,1 Department of Infectious Diseases and Immunology, Utrecht University, 3584 CL Utrecht, The Netherlands,2 WHO Collaborating Centre for Campylobacter/OIE Reference Laboratory for Campylobacteriosis Utrecht, The Netherlands3

Received 24 December 2008/ Accepted 3 April 2009

The presence and functionality of DNA repair mechanisms in Campylobacter jejuni are largely unknown. In silico analysis of the complete translated genome of C. jejuni NCTC 11168 suggests the presence of genes involved in methyl-directed mismatch repair (MMR), nucleotide excision repair, base excision repair (BER), and recombinational repair. To assess the functionality of these putative repair mechanisms in C. jejuni, mutS, uvrB, ung, and recA knockout mutants were constructed and analyzed for their ability to repair spontaneous point mutations, UV irradiation-induced DNA damage, and nicked DNA. Inactivation of the different putative DNA repair genes did not alter the spontaneous mutation frequency. Disruption of the UvrB and RecA orthologues, but not the putative MutS or Ung proteins, resulted in a significant reduction in viability after exposure to UV irradiation. Assays performed with uracil-containing plasmid DNA showed that the putative uracil-DNA glycosylase (Ung) protein, important for initiation of the BER pathway, is also functional in C. jejuni. Inactivation of recA also resulted in a loss of natural transformation. Overall, the data indicate that C. jejuni has multiple functional DNA repair systems that may protect against DNA damage and limit the generation of genetic diversity. On the other hand, the apparent absence of a functional MMR pathway may enhance the frequency of on-and-off switching of phase variable genes typical for C. jejuni and may contribute to the genetic heterogeneity of the C. jejuni population.

* Corresponding author. Mailing address: Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands. Phone: (31) 030 2534376. Fax: (31) 030 2533199. E-mail: J.Wagenaar{at}uu.nl

{triangledown} Published ahead of print on 17 April 2009.

{ddagger} Present address: Add2X Biosciences BV, 2333 AL Leiden, The Netherlands.

Journal of Bacteriology, June 2009, p. 3785-3793, Vol. 191, No. 12
0021-9193/09/$08.00+0     doi:10.1128/JB.01817-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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