Influence of BrpA on critical virulence attributes of Streptococcus mutans.
Streptococcus mutans, the primary etiological agent of human dental caries, has developed multiple mechanisms to colonize and form biofilms on the tooth surface. The brpA gene codes for a predicted surface-associated protein with apparent roles in biofilm formation, autolysis, and cell division. In this study, we used two models to further characterize the biofilm-forming characteristics of a BrpA-deficient mutant, strain TW14. Compared to those of the parent strain, UA159, TW14 formed long chains and sparse microcolonies on hydroxylapatite disks but failed to accumulate and form three-dimensional biofilms when grown on glucose as the carbohydrate source. The biofilm formation defect was also readily apparent by confocal laser scanning microscopy when flow cells were used to grow biofilms. When subjected to acid killing at pH 2.8 for 45 min, the survival rate of strain TW14 was more than 1 log lower than that of the wild-type strain. TW14 was at least 3 logs more susceptible to killing by 0.2% hydrogen peroxide than was UA159. The expression of more than 200 genes was found by microarray analysis to be altered in cells lacking BrpA (P < 0.01). These results suggest that the loss of BrpA can dramatically influence the transcriptome and significantly affects the regulation of acid and oxidative stress tolerance and biofilm formation in S. mutans, which are key virulence attributes of the organism.
Department of Oral Biology, College of Dentistry, University of Florida, Gainesville, FL 32610, USA.
Zezhang T
ZT
Wen
Henry V
HV
Baker
Robert A
RA
Burne
D000143:Acids / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D018441:Biofilms / Q000254:growth & development*; D017886:Durapatite; D015180:Electrophoresis, Gel, Two-Dimensional; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress; D020543:Proteome / Q000032:analysis / Q000302:isolation & purification; D013295:Streptococcus mutans / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity*; D014774:Virulence / Q000235:genetics / Q000502:physiology*
D000143:Acids; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; C454866:BrpA protein, Streptococcus mutans; D020543:Proteome; D017886:Durapatite