Department of Oral and Craniofacial Biology and Department of Microbiology, Immunology and Parasitology, LSU Health Sciences Center, New Orleans, LA, USA.
Mol Oral Microbiol. 2011 Feb;26(1):2-18. doi: 10.1111/j.2041-1014.2010.00581.x. Epub 2010 Nov 18.
We previously reported that LuxS in Streptococcus mutans is involved in stress tolerance and biofilm formation. In this study, flowcells and confocal laser scanning microscopy were used to further examine the effects of LuxS-deficiency on biofilm formation. Similar to the wild-type strain (UA159), a strain deficient in LuxS (TW26D) bound efficiently to the flowcells and formed microcolonies 4 h after inoculation. Unlike UA159, which accumulated and formed compact, evenly distributed biofilms after 28 h, TW26D showed only loose, sporadic, thin biofilms. DNA microarray analysis revealed alterations in transcription of more than 60 genes in TW26D biofilms by at least 1.5-fold (P < 0.001). Among the upregulated genes were those for sugar-specific enzymes II of the phosphotransferase (PTS) system and the atp operon, which codes for the proton-pumping F-ATPase. Of the downregulated genes, several encode proteins with putative functions in DNA repair. Mutation of selected genes caused severe defects in the ability of the mutants to tolerate low pH and oxidative stress. These results provide additional proof that LuxS-deficiency causes global alterations in the expression of genes central to biofilm formation and virulence of S. mutans, including those involved in energy metabolism, DNA repair and stress tolerance.
我们之前曾报道过变异链球菌中的 LuxS 参与了应激耐受和生物膜形成。在这项研究中,我们使用流槽和共聚焦激光扫描显微镜进一步研究了 LuxS 缺失对生物膜形成的影响。类似于野生型菌株(UA159),LuxS 缺失菌株(TW26D)能够有效地结合到流槽上,并在接种后 4 小时形成微菌落。与 UA159 不同,后者在 28 小时后积累并形成紧密、均匀分布的生物膜,而 TW26D 只显示出松散、零星、薄的生物膜。DNA 微阵列分析显示,TW26D 生物膜中超过 60 个基因的转录发生了至少 1.5 倍的改变(P<0.001)。上调的基因包括磷酸转移酶(PTS)系统中糖特异性酶 II 和 atp 操纵子的基因,该操纵子编码质子泵 F-ATP 酶。下调的基因中,有几个编码具有 DNA 修复功能的假定蛋白。选择基因的突变导致突变体在耐受低 pH 和氧化应激方面的能力严重缺陷。这些结果进一步证明了 LuxS 缺失导致了与变异链球菌生物膜形成和毒力相关的关键基因表达的全局改变,包括参与能量代谢、DNA 修复和应激耐受的基因。