Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-Sen University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, China.
Microbiologyopen. 2020 Sep;9(9):e1096. doi: 10.1002/mbo3.1096. Epub 2020 Jul 6.
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.
有证据表明，小型非编码 RNA（sRNA）参与了调控生物膜形成的复杂调控网络。很少有研究调查 sRNA 在变形链球菌（S. mutans）中的作用。本研究探讨了 sRNA 与 S. mutans 生物膜形成之间的关联。通过定量实时 PCR（qRT-PCR）鉴定了该细菌生物膜和浮游态中差异表达的 sRNA。使用共聚焦激光扫描显微镜研究了标准株 S. mutans（UA159，ATCC 700610）和 10 株临床分离株的生物膜形成特征。采用生物信息学分析预测和检查潜在的 sRNA 调控途径。结果表明，sRNA0426 与动态生物膜形成具有很强的正相关性。此外，sRNA0426 的表达与胞外多糖（EPS）的产生呈正相关。生物信息学分析表明，sRNA0426 参与生物膜形成的代谢途径，特别是碳代谢。选择了五个与 EPS 合成相关的靶 mRNA（GtfB、GtfC、GtfD、ComE 和 CcpA）进行进一步评估；其中三个 mRNA（GtfB、GtfC 和 CcpA）的表达水平与 sRNA0426 的表达水平呈正相关，一个（ComE）的表达水平呈负相关。总之，结果表明 sRNA0426 可能在 S. mutans 生物膜形成中发挥重要的积极作用，并为 S. mutans 生物膜调控网络提供新的见解。
