Yoshida Akihiro, Kuramitsu Howard K
Department of Oral Biology, State University of New York, Buffalo 14214, USA.
Appl Environ Microbiol. 2002 Dec;68(12):6283-91. doi: 10.1128/AEM.68.12.6283-6291.2002.
Streptococcus mutans has been strongly implicated as the principal etiological agent in dental caries. One of the important virulence properties of these organisms is their ability to form biofilms known as dental plaque on tooth surfaces. Since the roles of sucrose and glucosyltransferases in S. mutans biofilm formation have been well documented, we focused our attention on sucrose-independent factors. We have initially identified several mutants that appear to be defective in biofilm formation on abiotic surfaces by an insertional inactivation mutagenesis strategy applied to S. mutans. A total of 27 biofilm-defective mutants were isolated and analyzed in this study. From these mutants, three genes were identified. One of the mutants was defective in the Bacillus subtilis lytR homologue. Another of the biofilm-defective mutants isolated was a yulF homologue, which encodes a hypothetical protein of B. subtilis whose function in biofilm formation is unknown. The vast majority of the mutants were defective in the comB gene required for competence. We therefore have constructed and examined comACDE null mutants. These mutants were also found to be attenuated in biofilm formation. Biofilm formation by several other regulatory gene mutants were also characterized using an in vitro biofilm-forming assay. These results suggest that competence genes as well as the sgp and dgk genes may play important roles in S. mutans biofilm formation.
变形链球菌已被确认为龋齿的主要病原体。这些微生物的重要致病特性之一是它们能够在牙齿表面形成称为牙菌斑的生物膜。由于蔗糖和葡糖基转移酶在变形链球菌生物膜形成中的作用已有充分记录,我们将注意力集中在不依赖蔗糖的因素上。我们最初通过应用于变形链球菌的插入失活诱变策略,鉴定了几个似乎在非生物表面生物膜形成方面存在缺陷的突变体。本研究共分离并分析了27个生物膜缺陷突变体。从这些突变体中,鉴定出了三个基因。其中一个突变体在枯草芽孢杆菌lytR同源物中存在缺陷。另一个分离出的生物膜缺陷突变体是yulF同源物,它编码一种枯草芽孢杆菌的假定蛋白,其在生物膜形成中的功能尚不清楚。绝大多数突变体在感受态所需的comB基因中存在缺陷。因此,我们构建并检测了comACDE缺失突变体。这些突变体在生物膜形成方面也表现出减弱。还使用体外生物膜形成试验对其他几个调节基因突变体的生物膜形成进行了表征。这些结果表明,感受态基因以及sgp和dgk基因可能在变形链球菌生物膜形成中发挥重要作用。