Wang Bing, Kuramitsu Howard K
Department of Oral Biology, State University of New York at Buffalo, 3435 Main Street, Buffalo, New York 14214, USA.
Infect Immun. 2006 Aug;74(8):4581-9. doi: 10.1128/IAI.00001-06.
Exopolysaccharide synthesis, biofilm formation, and competence are important physiologic functions and virulence factors for Streptococcus mutans. In this study, we report the role of Frp, a transcriptional regulator, on the regulation of these traits crucial to pathogenesis. An Frp-deficient mutant showed decreased transcription of several genes important in virulence, including those encoding fructosyltransferase (Ftf), glucosyltransferase B (GtfB), and GtfC, by reverse transcription and quantitative real-time PCR. Expression of Ftf was decreased in the frp mutant, as assessed by Western blotting as well as by the activity assays. Frp deficiency also inhibited the production of GtfB in the presence of glucose and sucrose as well as the production of GtfC in the presence of glucose. As a consequence of the effects on GtfB and -C, sucrose-induced biofilm formation was decreased in the frp mutant. The expression of competence mediated by the competence-signaling peptide (CSP) system, as assessed by comC gene transcription, was attenuated in the frp mutant. As a result, the transformation efficiency was decreased in the frp mutant but was partially restored by adding synthetic CSP. Transcription of the frp gene was significantly increased in the frp mutant under all conditions tested, indicating that frp transcription is autoregulated. Furthermore, complementation of the frp gene in the frp mutant restored transcription of the affected genes to levels similar to those in the wild-type strain. These results suggest that Frp is a novel pleiotropic effector of multiple cellular functions and is involved in the modulation of exopolysaccharide synthesis, sucrose-dependent biofilm formation, and competence development.
胞外多糖合成、生物膜形成和感受态是变形链球菌重要的生理功能和毒力因子。在本研究中,我们报道了转录调节因子Frp在调控这些对发病机制至关重要的特性中的作用。通过逆转录和定量实时PCR分析,Frp缺陷型突变体中几个对毒力重要的基因转录水平降低,包括编码果糖基转移酶(Ftf)、葡糖基转移酶B(GtfB)和GtfC的基因。通过蛋白质免疫印迹和活性测定评估,Ftf在frp突变体中的表达降低。Frp缺陷还抑制了在葡萄糖和蔗糖存在下GtfB的产生以及在葡萄糖存在下GtfC的产生。由于对GtfB和GtfC的影响,frp突变体中蔗糖诱导的生物膜形成减少。通过comC基因转录评估,由感受态信号肽(CSP)系统介导的感受态表达在frp突变体中减弱。结果,frp突变体的转化效率降低,但通过添加合成CSP可部分恢复。在所有测试条件下,frp突变体中frp基因的转录显著增加,表明frp转录是自我调节的。此外,在frp突变体中互补frp基因可将受影响基因的转录恢复到与野生型菌株相似的水平。这些结果表明,Frp是多种细胞功能的新型多效效应因子,参与胞外多糖合成、蔗糖依赖性生物膜形成和感受态发育的调节。