Functional characterization of cell-wall-associated protein WapA in Streptococcus mutans.
Streptococcus mutans is known as a primary pathogen responsible for dental caries. One of the virulence factors of S. mutans in cariogenicity is its ability to attach to the tooth surface and form a biofilm. Several surface proteins have been shown to be involved in this process. A 29 kDa surface protein named wall-associated protein A (WapA, antigen A or antigen III), was previously used as a vaccine in animal studies for immunization against dental caries. However, the function of WapA in S. mutans is still not clear. This study characterized the function of WapA in cell surface structure and biofilm formation. Compared to the wild-type, the wapA mutant had much-reduced cell chain length, diminished cell-cell aggregation, altered cell surface ultrastructure, and unstructured biofilm architecture. Furthermore, in vivo force spectroscopy revealed that the cell surface of the wapA mutant was less sticky than that of the wild-type cells. More interestingly, these phenotypic differences diminished as sucrose concentration in the medium was increased to 0.5 %. Real-time RT-PCR analysis demonstrated that sucrose strongly repressed wapA gene expression in both planktonic and biofilm cells. These results suggest that the WapA protein plays an important structural role on the cell surface, which ultimately affects sucrose-independent cell-cell aggregation and biofilm architecture.
Zhu
Lin
L
School of Dentistry, University of California, Los Angeles, CA 90095, USA.
Kreth
Jens
J
School of Dentistry, University of California, Los Angeles, CA 90095, USA.
Cross
Sarah E
SE
Institute for Cell Mimetic Space Exploration, University of California, Los Angeles, CA 90095, USA.|Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, USA.
Gimzewski
James K
JK
California NanoSystems Institute, University of California, Los Angeles, CA 90095, USA.|Institute for Cell Mimetic Space Exploration, University of California, Los Angeles, CA 90095, USA.|Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, USA.
Shi
Wenyuan
W
Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA.|School of Dentistry, University of California, Los Angeles, CA 90095, USA.
Qi
Fengxia
F
School of Dentistry, University of California, Los Angeles, CA 90095, USA.
D000942:Antigens, Bacterial / Q000235:genetics / Q000502:physiology*; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D018441:Biofilms; D002462:Cell Membrane / Q000648:ultrastructure; D010641:Phenotype; D013295:Streptococcus mutans / Q000502:physiology* / Q000648:ultrastructure; D013395:Sucrose / Q000494:pharmacology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C081813:wapA protein, Bacillus subtilis; D013395:Sucrose