scholarly journals Trigger Factor in Streptococcus mutans Is Involved in Stress Tolerance, Competence Development, and Biofilm Formation

2005 ◽  
Vol 73 (1) ◽  
pp. 219-225 ◽  
Author(s):  
Zezhang T. Wen ◽  
Prashanth Suntharaligham ◽  
Dennis G. Cvitkovitch ◽  
Robert A. Burne
Keyword(s):  
Stress Tolerance ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Trigger Factor ◽  
Deficient Mutant ◽  
Carbohydrate Source ◽  
Oxidative Stresses ◽  
Parent Loss ◽  
Type Strains ◽  
And Oxidative Stress

ABSTRACT Trigger factor is a ribosome-associated peptidyl-prolyl cis/trans isomerase that is highly conserved in most bacteria. A gene, designated ropA, encoding an apparent trigger factor homologue, was identified in Streptococcus mutans, the primary etiological agent of human dental caries. Inactivation of ropA had no major impact on growth rate in planktonic cultures under the conditions tested, although the RopA-deficient mutant formed long chains in broth. Deficiency of RopA decreased tolerance to acid killing and to oxidative stresses induced by hydrogen peroxide and paraquat, and it reduced transformation efficiency about 200-fold. Addition of synthetic competence-stimulating peptide to the culture medium enhanced transformability of both the mutant and wild-type strains, although the ropA strain did not attain levels of competence observed for the parent. Loss of RopA decreased the capacity of S. mutans to form biofilms by over 80% when cultivated in glucose, but it increased biofilm formation by over 50% when sucrose was provided as the carbohydrate source. Western blot analysis revealed that the expression of glucosyltransferases B and D was lower in the RopA-deficient mutant. These results suggest that RopA is a key regulator of acid and oxidative stress tolerance, genetic competence, and biofilm formation, all critical virulence properties of S. mutans.

scholarly journals Streptococcus mutans Lacking sufCDSUB Is Viable, but Displays Major Defects in Growth, Stress Tolerance Responses and Biofilm Formation

2021 ◽  
Vol 12 ◽  
Author(s):  
Kassapa Ellepola ◽  
Xiaochang Huang ◽  
Ryan P. Riley ◽  
Jacob P. Bitoun ◽  
Zezhang Tom Wen
Keyword(s):  
Stress Tolerance ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Nitrosative Stress ◽  
Growth Stress ◽  
Low Ph ◽  
Luciferase Reporter ◽  
Deficient Mutant ◽  
Oxidative And Nitrosative Stress ◽  
Isoleucine Leucine

Streptococcus mutans appears to possess a sole iron-sulfur (Fe-S) cluster biosynthesis system encoded by the sufCDSUB cluster. This study was designed to examine the role of sufCDSUB in S. mutans physiology. Allelic exchange mutants deficient of the whole sufCDSUB cluster and in individual genes were constructed. Compared to the wild-type, UA159, the sufCDSUB-deficient mutant, Δsuf::kanr, had a significantly reduced growth rate, especially in medium with the absence of isoleucine, leucine or glutamate/glutamine, amino acids that require Fe-S clusters for biosynthesis and when grown with medium adjusted to pH 6.0 and under oxidative and nitrosative stress conditions. Relative to UA159, Δsuf::kanr had major defects in stress tolerance responses with reduced survival rate of > 2-logs following incubation at low pH environment or after hydrogen peroxide challenge. When compared to UA159, Δsuf::kanr tended to form aggregates in broth medium and accumulated significantly less biofilm. As shown by luciferase reporter fusion assays, the expression of sufCDSUB was elevated by > 5.4-fold when the reporter strain was transferred from iron sufficient medium to iron-limiting medium. Oxidative stress induced by methyl viologen increased sufCDSUB expression by > 2-fold, and incubation in a low pH environment led to reduction of sufCDSUB expression by > 7-fold. These results suggest that lacking of SufCDSUB in S. mutans causes major defects in various cellular processes of the deficient mutant, including growth, stress tolerance responses and biofilm formation. In addition, the viability of the deficient mutant also suggests that SUF, the sole Fe-S cluster machinery identified is non-essential in S. mutans, which is not known in any other bacterium lacking the NIF and/or ISC system. However, how the bacterium compensates the Fe-S deficiency and if any novel Fe-S assembly systems exist in this bacterium await further investigation.

scholarly journals Trk2 Potassium Transport System in Streptococcus mutans and Its Role in Potassium Homeostasis, Biofilm Formation, and Stress Tolerance

2016 ◽  
Vol 198 (7) ◽  
pp. 1087-1100 ◽  
Author(s):  
Gursonika Binepal ◽  
Kamal Gill ◽  
Paula Crowley ◽  
Martha Cordova ◽  
L. Jeannine Brady ◽  
...  
Keyword(s):  
Stress Tolerance ◽  
Streptococcus Mutans ◽  
Transport System ◽  
Biofilm Formation ◽  
Cellular Response ◽  
Pathogenic Bacteria ◽  
Transport Systems ◽  
Content Type ◽  
Growth And Survival ◽  
Dental Biofilm

ABSTRACTPotassium (K+) is the most abundant cation in the fluids of dental biofilm. The biochemical and biophysical functions of K+and a variety of K+transport systems have been studied for most pathogenic bacteria but not for oral pathogens. In this study, we establish the modes of K+acquisition inStreptococcus mutansand the importance of K+homeostasis for its virulence attributes. TheS. mutansgenome harbors four putative K+transport systems that included two Trk-like transporters (designated Trk1 and Trk2), one glutamate/K+cotransporter (GlnQHMP), and a channel-like K+transport system (Kch). Mutants lacking Trk2 had significantly impaired growth, acidogenicity, aciduricity, and biofilm formation. [K+] less than 5 mM eliminated biofilm formation inS. mutans. The functionality of the Trk2 system was confirmed by complementing anEscherichia coliTK2420 mutant strain, which resulted in significant K+accumulation, improved growth, and survival under stress. Taken together, these results suggest that Trk2 is the main facet of the K+-dependent cellular response ofS. mutansto environment stresses.IMPORTANCEBiofilm formation and stress tolerance are important virulence properties of caries-causingStreptococcus mutans. To limit these properties of this bacterium, it is imperative to understand its survival mechanisms. Potassium is the most abundant cation in dental plaque, the natural environment ofS. mutans. K+is known to function in stress tolerance, and bacteria have specialized mechanisms for its uptake. However, there are no reports to identify or characterize specific K+transporters inS. mutans. We identified the most important system for K+homeostasis and its role in the biofilm formation, stress tolerance, and growth. We also show the requirement of environmental K+for the activity of biofilm-forming enzymes, which explains why such high levels of K+would favor biofilm formation.

scholarly journals The Effects of Surface Roughness of Composite Resin on Biofilm Formation of Streptococcus mutans in the Presence of Saliva

2012 ◽  
Vol 37 (5) ◽  
pp. 532-539 ◽  
Author(s):  
JW Park ◽  
CW Song ◽  
JH Jung ◽  
SJ Ahn ◽  
JL Ferracane
Keyword(s):  
Surface Roughness ◽  
Silicon Carbide ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Composite Resin ◽  
Resin Composite ◽  
Fluid Phase ◽  
Confocal Laser ◽  
Carbohydrate Source ◽  
Effect Of Surface

SUMMARY The purpose of this study was to investigate the effects of surface roughness of resin composite on biofilm formation of Streptococcus mutans in the presence of saliva. To provide uniform surface roughness on composites, disks were prepared by curing composite against 400-grit silicon carbide paper (SR400), 800-grit silicon carbide paper (SR800), or a glass slide (SRGlass). The surface roughness was examined using confocal laser microscopy. For biofilm formation, S. mutans was grown for 24 hours with each disk in a biofilm medium with either glucose or sucrose in the presence of fluid-phase or surface-adsorbed saliva. The adherent bacteria were quantified via enumeration of the total viable counts of bacteria. Biofilms were examined using scanning electron microscopy. This study showed that SR400 had deeper and larger, but fewer depressions than SR800. Compared to SRGlass and SR800, biofilm formation was significantly increased on SR400. In addition, the differences in the effect of surface roughness on the amount of biofilm formation were not significantly influenced by either the presence of saliva or the carbohydrate source. Considering that similar differences in surface roughness were observed between SR400 and SR800 and between SR800 and SRGlass, this study suggests that surface topography (size and depth of depressions) may play a more important role than surface roughness in biofilm formation of S. mutans.

scholarly journals Inactivation of the ciaH Gene in Streptococcus mutans Diminishes Mutacin Production and Competence Development, Alters Sucrose-Dependent Biofilm Formation, and Reduces Stress Tolerance

2004 ◽  
Vol 72 (8) ◽  
pp. 4895-4899 ◽  
Author(s):  
Fengxia Qi ◽  
Justin Merritt ◽  
Renate Lux ◽  
Wenyuan Shi
Keyword(s):  
Stress Tolerance ◽  
Streptococcus Mutans ◽  
Histidine Kinase ◽  
Dental Plaque ◽  
Biofilm Formation ◽  
Competence Development ◽  
Clinical Isolates ◽  
Peptide Antibiotics ◽  
Kinase Gene

ABSTRACT Many clinical isolates of Streptococcus mutans produce peptide antibiotics called mutacins. Mutacin production may play an important role in the ecology of S. mutans in dental plaque. In this study, inactivation of a histidine kinase gene, ciaH, abolished mutacin production. Surprisingly, the same mutation also diminished competence development, stress tolerance, and sucrose-dependent biofilm formation.

scholarly journals A Hypothetical Protein of Streptococcus mutans Is Critical for Biofilm Formation

2005 ◽  
Vol 73 (5) ◽  
pp. 3147-3151 ◽  
Author(s):  
Thomas A. Brown ◽  
Sang-Joon Ahn ◽  
Roslyn N. Frank ◽  
Yi-Ywan M. Chen ◽  
José A. Lemos ◽  
...  
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Signal Sequence ◽  
Hypothetical Protein ◽  
Repeated Sequences ◽  
Carbohydrate Source ◽  
Novel Therapeutics ◽  
Potential Target ◽  
Conserved Regions

ABSTRACT Inactivation of the Smu0630 gene of Streptococcus mutans resulted in dramatic decreases in biofilm formation, regardless of the carbohydrate source. The Smu0630 protein contained numerous interesting features, including a possible signal sequence and two conserved regions of repeated sequences. Smu0630 may represent a potential target for novel therapeutics.

scholarly journals Role of Streptococcus gordoniiAmylase-Binding Protein A in Adhesion to Hydroxyapatite, Starch Metabolism, and Biofilm Formation

2001 ◽  
Vol 69 (11) ◽  
pp. 7046-7056 ◽  
Author(s):  
Jeffrey D. Rogers ◽  
Robert J. Palmer ◽  
Paul E. Kolenbrander ◽  
Frank A. Scannapieco
Keyword(s):  
Biofilm Formation ◽  
Binding Protein ◽  
Protein A ◽  
Scatchard Analysis ◽  
Wild Type ◽  
Salivary Amylase ◽  
Carbohydrate Source ◽  
Whole Saliva ◽  
Mutant Strains ◽  
Type Strains

ABSTRACT Interactions between bacteria and salivary components are thought to be important in the establishment and ecology of the oral microflora. α-Amylase, the predominant salivary enzyme in humans, binds to Streptococcus gordonii, a primary colonizer of the tooth. Previous studies have implicated this interaction in adhesion of the bacteria to salivary pellicles, catabolism of dietary starches, and biofilm formation. Amylase binding is mediated at least in part by the amylase-binding protein A (AbpA). To study the function of this protein, an erythromycin resistance determinant [erm(AM)] was inserted within the abpAgene of S. gordonii strains Challis and FAS4 by allelic exchange, resulting in abpA mutant strains Challis-E1 and FAS4-E1. Comparison of the wild-type and mutant strains did not reveal any significant differences in colony morphology, biochemical metabolic profiles, growth in complex or defined media, surface hydrophobicity, or coaggregation properties. Scatchard analysis of adhesion isotherms demonstrated that the wild-type strains adhered better to human parotid-saliva- and amylase-coated hydroxyapatite than did the AbpA mutants. In contrast, the mutant strains bound to whole-saliva-coated hydroxyapatite to a greater extent than did the wild-type strains. While the wild-type strains preincubated with purified salivary amylase grew well in defined medium with potato starch as the sole carbohydrate source, the AbpA mutants did not grow under the same conditions even after preincubation with amylase. In addition, the wild-type strain produced large microcolonies in a flow cell biofilm model, while the abpA mutant strains grew much more poorly and produced relatively small microcolonies. Taken together, these results suggest that AbpA ofS. gordonii functions as an adhesin to amylase-coated hydroxyapatite, in salivary-amylase-mediated catabolism of dietary starches and in human saliva-supported biofilm formation by S. gordonii.

scholarly journals Characteristics of Biofilm Formation by Streptococcus mutans in the Presence of Saliva

2008 ◽  
Vol 76 (9) ◽  
pp. 4259-4268 ◽  
Author(s):  
Sug-Joon Ahn ◽  
Sang-Joon Ahn ◽  
Zezhang T. Wen ◽  
L. Jeannine Brady ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Parental Strain ◽  
Biofilm Formation ◽  
Fluid Phase ◽  
Trigger Factor ◽  
Bacterial Aggregation ◽  
Whole Saliva ◽  
Tooth Surfaces ◽  
Biofilm Development ◽  
Induced Aggregation

ABSTRACT Interactions between salivary agglutinin and the adhesin P1 of Streptococcus mutans contribute to bacterial aggregation and mediate sucrose-independent adherence to tooth surfaces. We have examined biofilm formation by S. mutans UA159, and derivative strains carrying mutations affecting the localization or expression of P1, in the presence of fluid-phase or adsorbed saliva or salivary agglutinin preparations. Whole saliva- and salivary agglutinin-induced aggregation of S. mutans was adversely affected by the loss of P1 and sortase (SrtA) but not by the loss of trigger factor (RopA). Fluid-phase salivary agglutinin and, to a lesser extent, immobilized agglutinin inhibited biofilm development by S. mutans in the absence of sucrose, and whole saliva was more effective at decreasing biofilm formation than salivary agglutinin. Inhibition of biofilm development by salivary agglutinin was differently influenced by particular mutations, with the P1-deficient strain displaying a greater inhibition of biofilm development than the SrtA- or RopA-deficient strains. As expected, biofilm-forming capacities of all strains in the presence of salivary preparations were markedly enhanced in the presence of sucrose, although biofilm formation by the mutants was less efficient than that by the parental strain. Aeration strongly inhibited biofilm development, and the presence of salivary components did not restore biofilm formation in aerated conditions. The results disclose a potent ability of salivary constituents to moderate biofilm formation by S. mutans through P1-dependent and P1-independent pathways.

scholarly journals Effects of RelA on Key Virulence Properties of Planktonic and Biofilm Populations of Streptococcus mutans

2004 ◽  
Vol 72 (3) ◽  
pp. 1431-1440 ◽  
Author(s):  
José A. C. Lemos ◽  
Thomas A. Brown ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Type Strain ◽  
Wild Type Strain ◽  
Stringent Response ◽  
Strain Differences ◽  
Acid Tolerance ◽  
Normal Growth ◽  
Wild Type ◽  
Carbohydrate Source

ABSTRACT Streptococcus mutans is a biofilm-forming bacterium that is adapted to tolerate rapid and dramatic fluctuations in nutrient availability, carbohydrate source, and pH in its natural environment, the human oral cavity. Dissecting the pathways used to form stable biofilms and to tolerate environmental stress is central to understanding the virulence of this organism. Here, we investigated the role of the S. mutans relA gene, which codes for a guanosine tetraphosphate and guanosine pentaphosphate [(p)ppGpp] synthetase/hydrolase, in biofilm formation and acid tolerance. Two mutants in which relA was insertionally inactivated or replaced by an antibiotic resistance determinant were constructed. Under normal growth and stress conditions, the mutants grew slower than the wild-type strain, although the final yields were similar. The mutants, which were still able to accumulate (p)ppGpp after the induction of a stringent response, showed significant reductions in biofilm formation on microtiter plates or hydroxylapatite disks. There was no difference in the sensitivities to acid killing of the parent and relA strains grown in planktonic cultures. However, when cells were grown in biofilms, the mutants became more acid resistant and could lower the pH through glycolysis faster and to a greater extent than the wild-type strain. Differences in acid resistance were not correlated with increases in F-ATPase activity, although bacterial sugar:phosphotransferase activity was elevated in the mutants. Expression of the luxS gene was increased as much as fivefold in the relA mutants, suggesting a link between AI-2 quorum sensing and the stringent response.

scholarly journals Influence of BrpA on Critical Virulence Attributes of Streptococcus mutans

2006 ◽  
Vol 188 (8) ◽  
pp. 2983-2992 ◽  
Author(s):  
Zezhang T. Wen ◽  
Henry V. Baker ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Tooth Surface ◽  
Confocal Laser ◽  
Carbohydrate Source ◽  
Flow Cells ◽  
Forming Characteristics

ABSTRACT 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.

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