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Infection and Immunity, August 1999, p. 3909-3914, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Inactivation of the gbpA Gene of Streptococcus mutans Alters Structural and Functional Aspects of Plaque Biofilm Which Are Compensated by Recombination of the gtfB and gtfC Genes

Karsten R. O. Hazlett,dagger Joseph E. Mazurkiewicz, and Jeffrey A. Banas*

Department of Microbiology, Immunology, and Molecular Genetics, Albany Medical College, Albany, New York 12208

Received 5 March 1999/Returned for modification 4 May 1999/Accepted 27 May 1999

Inactivation of the gbpA gene of Streptococcus mutans increases virulence in a gnotobiotic rat model and also promotes in vivo accumulation of organisms in which gtfB and gtfC have recombined to reduce virulence (K. R. O. Hazlett, S. M. Michalek, and J. A. Banas, Infect. Immun. 66:2180-2185, 1998). These changes in virulence were hypothesized to result from changes in plaque structure. We have utilized an in vitro plaque model to test the hypothesis that the absence of GbpA alters S. mutans plaque structure and that the presence of gtfBC recombinant organisms within a gbpA background restores a wild-type (wt)-like plaque structure. When grown in the presence of sucrose within hydroxyapatite-coated wells, the wt S. mutans plaque consisted primarily of large aggregates which did not completely coat the hydroxyapatite surface, whereas the gbpA mutant plaque consisted of a uniform layer of smaller aggregates which almost entirely coated the hydroxyapatite. If 25% of the gbpA mutants used as inoculum were also gtfBC recombinants (gbpA/25%gtfBC), a wt-like plaque was formed. These changes in plaque structure correlated with differences in susceptibility to ampicillin; gbpA plaque organisms were more susceptible than organisms in either the wt or gbpA/25%gtfBC plaques. These data allow the conclusion that GbpA contributes to S. mutans plaque biofilm development. Since the changes in plaque structure detailed in this report correlate well with previously observed changes in virulence, it seems likely that S. mutans biofilm structure influences virulence. A potential model for this influence, which can account for the gtfBC recombination compensating gbpA inactivation, is that the ratio of glucan to glucan-binding protein is a critical factor in plaque development.

* Corresponding author. Mailing address: Department of Microbiology, Immunology, and Molecular Genetics, Albany Medical College, Albany, NY 12208. Phone: (518) 262-6286. Fax: (518) 262-5748. E-mail: Jeff_Banas{at}ccgateway.amc.edu.

dagger Present address: Center for Microbial Pathogenesis, University of Connecticut Health Center, Farmington, CT 06032.

Infection and Immunity, August 1999, p. 3909-3914, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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