scholarly journals Analysis of an Agmatine Deiminase Gene Cluster in Streptococcus mutans UA159

2004 ◽  
Vol 186 (6) ◽  
pp. 1902-1904 ◽  
Author(s):  
Ann R. Griswold ◽  
Yi-Ywan M. Chen ◽  
Robert A. Burne
Keyword(s):  
Gene Cluster ◽  
Streptococcus Mutans ◽  
Catabolite Repression ◽  
Acid Tolerance ◽  
Low Ph ◽  
Agmatine Deiminase ◽  
Cariogenic Bacterium

ABSTRACT An operon encoding enzymes of the agmatine deiminase system (AgDS) has been identified in the cariogenic bacterium Streptococcus mutans UA159. The AgDS is regulated by agmatine induction and carbohydrate catabolite repression. Ammonia is produced from agmatine at low pH, suggesting that the AgDS could augment acid tolerance.

scholarly journals Regulation and Physiologic Significance of the Agmatine Deiminase System of Streptococcus mutans UA159

2006 ◽  
Vol 188 (3) ◽  
pp. 834-841 ◽  
Author(s):  
Ann R. Griswold ◽  
Max Jameson-Lee ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Transcription Initiation ◽  
Acid Tolerance ◽  
Acid Resistance ◽  
Initiation Site ◽  
Low Ph ◽  
Pathogenic Potential ◽  
Genome Database ◽  
Reading Frame ◽  
Agmatine Deiminase

ABSTRACT We previously demonstrated that Streptococcus mutans expresses a functional agmatine deiminase system (AgDS) encoded by the agmatine-inducible aguBDAC operon (A. R. Griswold, Y. Y. Chen, and R. A. Burne, J. Bacteriol. 186:1902-1904, 2004). The AgDS yields ammonia, CO2, and ATP while converting agmatine to putrescine and is proposed to augment the acid resistance properties and pathogenic potential of S. mutans. To initiate a study of agu gene regulation, the aguB transcription initiation site was identified by primer extension and a putative σ70-like promoter was mapped 5′ to aguB. Analysis of the genome database revealed an open reading frame (SMU.261c) encoding a putative transcriptional regulator located 239 bases upstream of aguB. Inactivation of SMU.261c decreased AgD activity by sevenfold and eliminated agmatine induction. AgD was also found to be induced by certain environmental stresses, including low pH and heat, implying that the AgDS may also be a part of a general stress response pathway of this organism. Interestingly, an AgDS-deficient strain was unable to grow in the presence of 20 mM agmatine, suggesting that the AgDS converts a growth-inhibitory substance into products that can enhance acid tolerance and contribute to the competitive fitness of the organism at low pH. The capacity to detoxify and catabolize agmatine is likely to have major ramifications on oral biofilm ecology.

scholarly journals AguR Is Required for Induction of the Streptococcus mutans Agmatine Deiminase System by Low pH and Agmatine

2009 ◽  
Vol 75 (9) ◽  
pp. 2629-2637 ◽  
Author(s):  
Yaling Liu ◽  
Lin Zeng ◽  
Robert A. Burne
Keyword(s):  
Gene Expression ◽  
Dna Binding ◽  
Streptococcus Mutans ◽  
Low Ph ◽  
Binding Domain ◽  
Negative Regulator ◽  
Dna Binding Domain ◽  
Mutant Proteins ◽  
Acidic Conditions ◽  
Agmatine Deiminase

ABSTRACT Acidic conditions and the presence of exogenous agmatine are required to achieve maximal expression of the agmatine deiminase system (AgDS) of Streptococcus mutans. Here we demonstrate that the transcriptional activator of the AgDS, AguR, is required for the responses to agmatine and to low pH. Linker scanning mutagenesis was used to create a panel of mutated aguR genes that were utilized to complement an aguR deletion mutant of S. mutans. The level of production of the mutant proteins was shown to be comparable to that of the wild-type AguR protein. Mutations in the predicted DNA binding domain of AguR eliminated activation of the agu operon. Insertions into the region connecting the DNA binding domain to the predicted extracellular and transmembrane domains were well tolerated. In contrast, a variety of mutants were isolated that had a diminished capacity to respond to low pH but retained the ability to activate AgDS gene expression in response to agmatine, and vice versa. Also, a number of mutants were unable to respond to either agmatine or low pH. AguD, which is a predicted agmatine-putrescine antiporter, was found to be a negative regulator of AgDS gene expression in the absence of exogenous agmatine but was not required for low-pH induction of the AgDS genes. This study reveals that the control of AgDS gene expression by both agmatine and low pH is coordinated through the AguR protein and begins to identify domains of the protein involved in sensing and signaling.

scholarly journals Multiple Two-Component Systems of Streptococcus mutans Regulate Agmatine Deiminase Gene Expression and Stress Tolerance

2009 ◽  
Vol 191 (23) ◽  
pp. 7363-7366 ◽  
Author(s):  
Yaling Liu ◽  
Robert A. Burne
Keyword(s):  
Gene Expression ◽  
Stress Tolerance ◽  
Streptococcus Mutans ◽  
Thermal Stresses ◽  
Low Ph ◽  
Component Systems ◽  
Two Component ◽  
Two Component Systems ◽  
Agmatine Deiminase

ABSTRACT Induction of the agmatine deiminase system (AgDS) of Streptococcus mutans requires agmatine and is optimal at low pH. We show here that the VicRK, ComDE, and CiaRH two-component systems influence AgDS gene expression in response to acidic and thermal stresses.

Stress-responsive proteins are upregulated in Streptococcus mutans during acid tolerance

Microbiology ◽  
2004 ◽  
Vol 150 (5) ◽  
pp. 1339-1351 ◽  
Author(s):  
Alice C. L. Len ◽  
Derek W. S. Harty ◽  
Nicholas A. Jacques
Keyword(s):  
Streptococcus Mutans ◽  
Elongation Factor ◽  
Acid Tolerance ◽  
Proteome Analysis ◽  
Low Ph ◽  
Lactobacillus Helveticus ◽  
Oral Streptococci ◽  
Flight Mass Spectrometry ◽  
Future Assessment ◽  
Acid Tolerant

Streptococcus mutans is an important pathogen in the initiation of dental caries as the bacterium remains metabolically active when the environment becomes acidic. The mechanisms underlying this ability to survive and proliferate at low pH remain an area of intense investigation. Differential two-dimensional electrophoretic proteome analysis of S. mutans grown at steady state in continuous culture at pH 7·0 or pH 5·0 enabled the resolution of 199 cellular and extracellular protein spots with altered levels of expression. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry identified 167 of these protein spots. Sixty-one were associated with stress-responsive pathways involved in DNA replication, transcription, translation, protein folding and proteolysis. The 61 protein spots represented isoforms or cleavage products of 30 different proteins, of which 25 were either upregulated or uniquely expressed during acid-tolerant growth at pH 5·0. Among the unique and upregulated proteins were five that have not been previously identified as being associated with acid tolerance in S. mutans and/or which have not been studied in any detail in oral streptococci. These were the single-stranded DNA-binding protein, Ssb, the transcription elongation factor, GreA, the RNA exonuclease, polyribonucleotide nucleotidyltransferase (PnpA), and two proteinases, the ATP-binding subunit, ClpL, of the Clp family of proteinases and a proteinase encoded by the pep gene family with properties similar to the dipeptidase, PepD, of Lactobacillus helveticus. The identification of these and other differentially expressed proteins associated with an acid-tolerant-growth phenotype provides new information on targets for mutagenic studies that will allow the future assessment of their physiological significance in the survival and proliferation of S. mutans in low pH environments.

scholarly journals Regulation and Physiological Significance of ClpC and ClpP in Streptococcus mutans

2002 ◽  
Vol 184 (22) ◽  
pp. 6357-6366 ◽  
Author(s):  
José A. C. Lemos ◽  
Robert A. Burne
Keyword(s):  
Gene Expression ◽  
Streptococcus Mutans ◽  
Stress Responses ◽  
Expression Patterns ◽  
Acid Tolerance ◽  
Low Ph ◽  
Two Dimensional ◽  
Wild Type ◽  
Protein Gels

ABSTRACT Tolerance of environmental stress, especially low pH, by Streptococcus mutans is central to the virulence of this organism. The Clp ATPases are implicated in the tolerance of, and regulation of the response to, stresses by virtue of their protein reactivation and remodeling activities and their capacity to target misfolded proteins for degradation by the ClpP peptidase. The purpose of this study was to dissect the role of selected clp genes in the stress responses of S. mutans, with a particular focus on acid tolerance and adaptation. Homologues of the clpB, clpC, clpE, clpL, clpX, and clpP genes were identified in the S. mutans genome. The expression of clpC and clpP, which were chosen as the focus of this study, was induced at low pH and at growth above 40°C. Inactivation of ctsR, the first of two genes in the clpC operon, demonstrated that CtsR acts as a repressor of clp and groES-EL gene expression. Strains lacking ClpP, but not strains lacking ClpC, were impaired in their ability to grow under stress-inducing conditions, formed long chains, aggregated in culture, had reduced genetic transformation efficiencies, and had a reduced capacity to form biofilms. Comparison of two-dimensional protein gels from wild-type cells and the ctsR and clpP mutants revealed many changes in the protein expression patterns. In particular, in the clpP mutant, there was an increased production of GroESL and DnaK, suggesting that cells were stressed, probably due to the accumulation of denatured proteins.

scholarly journals Cell Density Modulates Acid Adaptation in Streptococcus mutans: Implications for Survival in Biofilms

2001 ◽  
Vol 183 (23) ◽  
pp. 6875-6884 ◽  
Author(s):  
Yung-Hua Li ◽  
Michael N. Hanna ◽  
Gunnel Svensäter ◽  
Richard P. Ellen ◽  
Dennis G. Cvitkovitch
Keyword(s):  
Streptococcus Mutans ◽  
Cell Density ◽  
Culture Filtrate ◽  
High Cell Density ◽  
Acid Tolerance ◽  
Low Ph ◽  
Growth Mode ◽  
High Cell ◽  
Biofilm Growth ◽  
Acid Adaptation

ABSTRACT Streptococcus mutans normally colonizes dental biofilms and is regularly exposed to continual cycles of acidic pH during ingestion of fermentable dietary carbohydrates. The ability ofS. mutans to survive at low pH is an important virulence factor in the pathogenesis of dental caries. Despite a few studies of the acid adaptation mechanism of this organism, little work has focused on the acid tolerance of S. mutans growing in high-cell-density biofilms. It is unknown whether biofilm growth mode or high cell density affects acid adaptation by S. mutans. This study was initiated to examine the acid tolerance response (ATR) of S. mutans biofilm cells and to determine the effect of cell density on the induction of acid adaptation. S. mutans BM71 cells were first grown in broth cultures to examine acid adaptation associated with growth phase, cell density, carbon starvation, and induction by culture filtrates. The cells were also grown in a chemostat-based biofilm fermentor for biofilm formation. Adaptation of biofilm cells to low pH was established in the chemostat by the acid generated from excess glucose metabolism, followed by a pH 3.5 acid shock for 3 h. Both biofilm and planktonic cells were removed to assay percentages of survival. The results showed that S. mutans BM71 exhibited a log-phase ATR induced by low pH and a stationary-phase acid resistance induced by carbon starvation. Cell density was found to modulate acid adaptation in S. mutans log-phase cells, since pre-adapted cells at a higher cell density or from a dense biofilm displayed significantly higher resistance to the killing pH than the cells at a lower cell density. The log-phase ATR could also be induced by a neutralized culture filtrate collected from a low-pH culture, suggesting that the culture filtrate contained an extracellular induction component(s) involved in acid adaptation in S. mutans. Heat or proteinase treatment abolished the induction by the culture filtrate. The results also showed that mutants defective in thecomC, -D, or -E genes, which encode a quorum sensing system essential for cell density-dependent induction of genetic competence, had a diminished log-phase ATR. Addition of synthetic competence stimulating peptide (CSP) to the comC mutant restored the ATR. This study demonstrated that cell density and biofilm growth mode modulated acid adaptation in S. mutans, suggesting that optimal development of acid adaptation in this organism involves both low pH induction and cell-cell communication.

scholarly journals Physiologic Effects of Forced Down-Regulation of dnaK and groEL Expression in Streptococcus mutans

2006 ◽  
Vol 189 (5) ◽  
pp. 1582-1588 ◽  
Author(s):  
José A. Lemos ◽  
Yaima Luzardo ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Parent Strain ◽  
Type Strain ◽  
Wild Type Strain ◽  
Acid Tolerance ◽  
Low Ph ◽  
Broth Culture ◽  
Wild Type ◽  
Compensatory Mutations

ABSTRACT Strains of Streptococcus mutans lacking DnaK or GroEL appear not to be isolable. To better distinguish the roles played by these chaperones/chaperonins in the physiology of S. mutans, we created a knockdown strategy to lower the levels of DnaK by over 95% in strain SM12 and the level of GroEL about 80% in strain SM13. Interestingly, GroEL levels were approximately twofold higher in SM12 than in the parent strain, but the levels of DnaK were not altered in the GroEL knockdown strain. Both SM12 and SM13 grew slower than the parent strain, had a strong tendency to aggregate in broth culture, and showed major changes in their proteomes. Compared with the wild-type strain, SM12 and SM13 had impaired biofilm-forming capacities when grown in the presence of glucose. The SM12 strain was impaired in its capacity to grow at 44°C or at pH 5.0 and was more susceptible to H2O2, whereas SM13 behaved like the wild-type strain under these conditions. Phenotypical reversions were noted for both mutants when cells were grown in continuous culture at a low pH, suggesting the occurrence of compensatory mutations. These results demonstrate that DnaK and GroEL differentially affect the expression of key virulence traits, including biofilm formation and acid tolerance, and support that these chaperones have evolved to accommodate unique roles in the context of this organism and its niche.

scholarly journals Modification of Gene Expression and Virulence Traits in Streptococcus mutans in Response to Carbohydrate Availability

2013 ◽  
Vol 80 (3) ◽  
pp. 972-985 ◽  
Author(s):  
Zachary D. Moye ◽  
Lin Zeng ◽  
Robert A. Burne
Keyword(s):  
Streptococcus Mutans ◽  
Catabolite Repression ◽  
Acid Tolerance ◽  
Phosphotransferase System ◽  
Content Type ◽  
Carbohydrate Availability ◽  
Biochemical Pathways ◽  
Profound Influence ◽  
Excess Glucose ◽  
Phenotypic Responses

ABSTRACTThe genetic and phenotypic responses ofStreptococcus mutans, an organism that is strongly associated with the development of dental caries, to changes in carbohydrate availability were investigated.S. mutansUA159 or a derivative of UA159 lacking ManL, which is the EIIAB component (EIIABMan) of a glucose/mannose permease of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) and a dominant effector of catabolite repression, was grown in continuous culture to steady state under conditions of excess (100 mM) or limiting (10 mM) glucose. Microarrays using RNA fromS. mutansUA159 revealed that 174 genes were differentially expressed in response to changes in carbohydrate availability (P< 0.001). Glucose-limited cells possessed higher PTS activity, could acidify the environment more rapidly and to a greater extent, and produced more ManL protein than cultures grown with excess glucose. Loss of ManL adversely affected carbohydrate transport and acid tolerance. Comparison of the histidine protein (HPr) inS. mutansUA159 and themanLdeletion strain indicated that the differences in the behaviors of the strains were not due to major differences in HPr pools or HPr phosphorylation status. Therefore, carbohydrate availability alone can dramatically influence the expression of physiologic and biochemical pathways that contribute directly to the virulence ofS. mutans, and ManL has a profound influence on this behavior.

scholarly journals Acid tolerance in early colonizers of oral biofilms

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Gabriella Boisen ◽  
Julia R. Davies ◽  
Jessica Neilands
Keyword(s):  
Acid Tolerance ◽  
Low Ph ◽  
Salivary Proteins ◽  
Confocal Scanning Laser Microscopy ◽  
Planktonic Cells ◽  
Tolerance Development ◽  
Salivary Pellicle ◽  
Oral Biofilms ◽  
Acid Tolerant

Abstract Background In caries, low pH drives selection and enrichment of acidogenic and aciduric bacteria in oral biofilms, and development of acid tolerance in early colonizers is thought to play a key role in this shift. Since previous studies have focussed on planktonic cells, the effect of biofilm growth as well as the role of a salivary pellicle on this process is largely unknown. We explored acid tolerance and acid tolerance response (ATR) induction in biofilm cells of both clinical and laboratory strains of three oral streptococcal species (Streptococcus gordonii, Streptococcus oralis and Streptococcus mutans) as well as two oral species of Actinomyces (A. naeslundii and A. odontolyticus) and examined the role of salivary proteins in acid tolerance development. Methods Biofilms were formed on surfaces in Ibidi® mini flow cells with or without a coating of salivary proteins and acid tolerance assessed by exposing them to a challenge known to kill non-acid tolerant cells (pH 3.5 for 30 min) followed by staining with LIVE/DEAD BacLight and confocal scanning laser microscopy. The ability to induce an ATR was assessed by exposing the biofilms to an adaptation pH (pH 5.5) for 2 hours prior to the low pH challenge. Results Biofilm formation significantly increased acid tolerance in all the clinical streptococcal strains (P < 0.05) whereas the laboratory strains varied in their response. In biofilms, S. oralis was much more acid tolerant than S. gordonii or S. mutans. A. naeslundii showed a significant increase in acid tolerance in biofilms compared to planktonic cells (P < 0.001) which was not seen for A. odontolyticus. All strains except S. oralis induced an ATR after pre-exposure to pH 5.5 (P < 0.05). The presence of a salivary pellicle enhanced both acid tolerance development and ATR induction in S. gordonii biofilms (P < 0.05) but did not affect the other bacteria to the same extent. Conclusions These findings suggest that factors such as surface contact, the presence of a salivary pellicle and sensing of environmental pH can contribute to the development of high levels of acid tolerance amongst early colonizers in oral biofilms which may be important in the initiation of caries.

Sign in / Sign up

Export Citation Format

Share Document