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Small noncoding RNA sRNA0426 is involved in regulating biofilm formation in Streptococcus mutans.

ABSTRACT: Evidence suggests that small noncoding RNAs (sRNAs) are involved in the complex regulatory networks governing biofilm formation. Few studies have investigated the role of sRNAs in Streptococcus mutans (S. mutans). In the present study, the association between sRNA and biofilm formation in S. mutans was explored. sRNAs that are differentially expressed in the biofilm and planktonic states of this bacterium were identified by quantitative real-time PCR (qRT-PCR). Confocal laser scanning microscopy was used to investigate the characteristics of biofilm formation in a standard strain of S. mutans (UA159, ATCC 700610) and ten clinical strains. Bioinformatics analyses were employed to predict and examine potential sRNA regulatory pathways. The results showed that sRNA0426 has a strong positive relationship with dynamic biofilm formation. Moreover, sRNA0426 expression was positively correlated with exopolysaccharide (EPS) production. Bioinformatics analyses showed that sRNA0426 is involved in biofilm formation such as metabolic pathways, especially carbon metabolism. Five target mRNAs (GtfB, GtfC, GtfD, ComE, and CcpA) involved in the synthesis of EPS were selected for further evaluation; the expression levels of three of these mRNAs (GtfB, GtfC, and CcpA) were positively correlated with sRNA0426 expression levels, and the expression level of one (ComE) was negatively correlated. In conclusion, the results suggested that sRNA0426 may play an important and positive role in the biofilm formation of S. mutans and provide novel insight into the S. mutans biofilm regulatory network.

SUBMITTER: Yin L

PROVIDER: S-EPMC7521000 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Small noncoding RNA sRNA0426 is involved in regulating biofilm formation in Streptococcus mutans.

Yin Luoping L Zhu Wenhui W Chen Dongru D Zhou Yan Y Lin Huancai H

MicrobiologyOpen 20200706 9

Evidence suggests that small noncoding RNAs (sRNAs) are involved in the complex regulatory networks governing biofilm formation. Few studies have investigated the role of sRNAs in Streptococcus mutans (S. mutans). In the present study, the association between sRNA and biofilm formation in S. mutans was explored. sRNAs that are differentially expressed in the biofilm and planktonic states of this bacterium were identified by quantitative real-time PCR (qRT-PCR). Confocal laser scanning microscopy ...[more]

PMID: 32633012

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Project description:The abilities of Streptococcus mutans to form biofilms and to survive acidic pH are regarded as two important virulence determinants in the pathogenesis of dental caries. Environmental stimuli are thought to regulate the expression of several genes associated with virulence factors through the activity of two-component signal transduction systems. Yet, little is known of the involvement of these systems in the physiology and pathogenicity of S. mutans. In this study, we describe a two-component regulatory system and its involvement in biofilm formation and acid resistance in S. mutans. By searching the S. mutans genome database with tblastn with the HK03 and RR03 protein sequences from S. pneumoniae as queries, we identified two genes, designated hk11 and rr11, that encode a putative histidine kinase and its cognate response regulator. To gain insight into their function, a PCR-mediated allelic-exchange mutagenesis strategy was used to create the hk11 (Em(r)) and rr11 (Em(r)) deletion mutants from S. mutans wild-type NG8 named SMHK11 and SMRR11, respectively. The mutants were examined for their growth rates, genetic competence, ability to form biofilms, and resistance to low-pH challenge. The results showed that deletion of hk11 or rr11 resulted in defects in biofilm formation and resistance to acidic pH. Both mutants formed biofilms with reduced biomass (50 to 70% of the density of the parent strain). Scanning electron microscopy revealed that the biofilms formed by the mutants had sponge-like architecture with what appeared to be large gaps that resembled water channel-like structures. The mutant biofilms were composed of longer chains of cells than those of the parent biofilm. Deletion of hk11 also resulted in greatly diminished resistance to low pH, although we did not observe the same effect when rr11 was deleted. Genetic competence was not affected in either mutant. The results suggested that the gene product of hk11 in S. mutans might act as a pH sensor that could cross talk with one or more response regulators. We conclude that the two-component signal transduction system encoded by hk11 and rr11 represents a new regulatory system involved in biofilm formation and acid resistance in S. mutans.

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Small noncoding RNA sRNA0426 is involved in regulating biofilm formation in Streptococcus mutans.

Publications

Small noncoding RNA sRNA0426 is involved in regulating biofilm formation in Streptococcus mutans.

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