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Journal of Bacteriology, June 2006, p. 3813-3825, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.01845-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Functional Analysis of Glucan Binding Protein B from Streptococcus mutans

Renata O. Mattos-Graner ,1,#,{dagger} Kristen A. Porter,2,# Daniel J. Smith,1 Yumiko Hosogi,2 and Margaret J. Duncan2*

Department of Immunology,1 Department of Molecular Genetics, The Forsyth Institute, Boston, Massachusetts 021152

Received 2 December 2005/ Accepted 16 March 2006

Mutans streptococci are major etiological agents of dental caries, and several of their secreted products contribute to bacterial accumulation on teeth. Of these, Streptococcus mutans glucan binding protein B (GbpB) is a novel, immunologically dominant protein. Its biological function is unclear, although GbpB shares homology with a putative peptidoglycan hydrolase from S. agalactiae and S. pneumoniae, indicative of a role in murein biosynthesis. To determine the cellular function of GbpB, we used several approaches to inactivate the gene, analyze its expression, and identify interacting proteins. None of the transformants analyzed were true gbpB mutants, since they all contained both disrupted and wild-type gene copies, and expression of functional GbpB was always conserved. Thus, the inability to obtain viable gbpB null mutants supports the notion that gbpB is an essential gene. Northern blot and real-time PCR analyses suggested that induction of gbpB expression in response to stress was a strain-dependent phenomenon. Proteins that interacted with GbpB were identified in pull-down and coimmunoprecipitation assays, and these data suggest that GbpB interacts with ribosomal protein L7/L12, possibly as part of a protein complex involved in peptidoglycan synthesis and cell division.

* Corresponding author. Mailing address: Department of Molecular Genetics, The Forsyth Institute, 140 Fenway, Boston, MA 02115. Phone: (617) 262-5200, ext. 8344. Fax: (617) 262-4021. E-mail: mduncan{at}forsyth.org.

# These authors made equal contributions to this work.

{dagger} Present address: Department of Microbiology and Immunology, Piracicaba School of Dentistry, University of Campinas, Sao Paulo, Brazil.

Journal of Bacteriology, June 2006, p. 3813-3825, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.01845-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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