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Infection and Immunity, August 2001, p. 5037-5045, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.5037-5045.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Tandem Repeat Deletion in the Alpha C Protein of Group B Streptococcus Is recA Independent

Karen M. Puopolo,1,2,3,* Susan K. Hollingshead,4 Vincent J. Carey,1,5 and Lawrence C. Madoff1,6

Channing Laboratory1 and Division of Infectious Diseases,6 Department of Medicine, and Department of Newborn Medicine,2 Brigham and Women's Hospital, and Division of Newborn Medicine, Children's Hospital,3 Harvard Medical School, and Department of Biostatistics, Harvard School of Public Health,5 Boston, Massachusetts, and Department of Microbiology, University of Alabama, Birmingham, Alabama4

Received 12 March 2001/Returned for modification 7 May 2001/Accepted 11 May 2001

Group B streptococci (GBS) contain a family of protective surface proteins characterized by variable numbers of repeating units within the proteins. The prototype alpha C protein of GBS from the type Ia/C strain A909 contains a series of nine identical 246-bp tandem repeat units. We have previously shown that deletions in the tandem repeat region of the alpha C protein affect both the immunogenicity and protective efficacy of the protein in animal models, and these deletions may serve as a virulence mechanism in GBS. The molecular mechanism of tandem repeat deletion is unknown. To determine whether RecA-mediated homologous recombination is involved in this process, we identified, cloned, and sequenced the recA gene homologue from GBS. A strain of GBS with recA deleted, A909Delta recA, was constructed by insertional inactivation in the recA locus. A909Delta recA demonstrated significant sensitivity to UV light, and the 50% lethal dose of the mutant strain in a mouse intraperitoneal model of sepsis was 20-fold higher than that of the parent strain. The spontaneous rate of tandem repeat deletion in the alpha C protein in vitro, as well as in our mouse model of immune infection, was studied using A909Delta recA. We report that tandem repeat deletion in the alpha C protein does occur in the absence of a functional recA gene both in vitro and in vivo, indicating that tandem repeat deletion in GBS occurs by a recA-independent recombinatorial pathway.

* Corresponding author. Mailing address: Channing Laboratory, 181 Longwood Ave., Boston, MA 02115. Phone: (617) 525-2681. Fax: (617) 731-1541. E-mail: kpuopolo{at}rics.bwh.harvard.edu.

Infection and Immunity, August 2001, p. 5037-5045, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.5037-5045.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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