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Infection and Immunity, October 2000, p. 5663-5667, Vol. 68, No. 10
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Campylobacter fetus sap Inversion Occurs in the Absence of RecA Function

Kevin C. Ray,1 Zheng-Chao Tu,1 Rosemary Grogono-Thomas,2 Diane G. Newell,3 Stuart A. Thompson,4 and Martin J. Blaser1,*

Vanderbilt University School of Medicine and VA Medical Center, Nashville, Tennessee1; Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, Georgia4; and Department of Farm Animal and Equine Medicine and Surgery, Royal Veterinary College, Potters Bar, Hertfordshire,2 and Veterinary Laboratories Agency (Weybridge), New Haw, Surrey,3 United Kingdom

Received 3 April 2000/Returned for modification 27 May 2000/Accepted 4 July 2000

Phase variation of Campylobacter fetus surface layer proteins (SLPs) occurs by inversion of a 6.2-kb DNA segment containing the unique sap promoter, permitting expression of a single SLP-encoding gene. Previous work has shown that the C. fetus sap inversion system is RecA dependent. When we challenged a pregnant ewe with a recA mutant of wild-type C. fetus (strain 97-211) that expressed the 97-kDa SLP, 15 of the 16 ovine-passaged isolates expressed the 97-kDa protein. However, one strain (97-209) expressed a 127-kDa SLP, suggesting that chromosomal rearrangement may have occurred to enable SLP switching. Lack of RecA function in strains 97-211 and 97-209 was confirmed by their sensitivity to the DNA-damaging agent methyl methanesulfonate. Southern hybridization and PCR of these strains indicated that the aphA insertion into recA was stably present. However, Southern hybridizations demonstrated that in strain 97-209 inversion had occurred in the sap locus. PCR data confirmed inversion of the 6.2-kb DNA element and indicated that in these recA mutants the sap inversion frequency is reduced by 2 to 3 log10 units compared to that in the wild type. Thus, although the major sap inversion pathway in C. fetus is RecA dependent, alternative lower-frequency, RecA-independent inversion mechanisms exist.

* Corresponding author. Mailing address: Department of Medicine, New York University Medical Center, 550 First Ave., New York, NY 10016. Phone: (212) 263-6394. Fax: (212) 263-7700. E-mail: martin.blaser{at}med.nyu.edu.

Infection and Immunity, October 2000, p. 5663-5667, Vol. 68, No. 10
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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