He Yiping, Frye Jonathan G, Strobaugh Terence P, Chen Chin-Yi
Microbial Biophysics and Residue Chemistry Research Unit, ERRC-ARS-USDA, Wyndmoor, Pennsylvania 19038, USA.
Foodborne Pathog Dis. 2008 Aug;5(4):399-415. doi: 10.1089/fpd.2008.0106.
Autoinducer-2 (AI-2) is a quorum-sensing signal molecule that controls a variety of cellular activities in response to cell density in both gram-negative and gram-positive bacteria. The production of AI-2 is dependent upon LuxS, the last enzyme in the AI-2 biosynthesis pathway. For this study, we constructed a luxS null mutation (Delta luxS) in Campylobacter jejuni strain 81-176, and showed that it abolished AI-2 production. The Delta luxS mutant had a longer doubling time in Mueller-Hinton (MH) broth and reduced swarming on MH soft agar at 37 degrees C compared to the wild type (wt), whereas growth rate or swarming at 42 degrees C was not affected. The Delta luxS mutant was also more sensitive to hydrogen peroxide (H(2)O(2)) and cumene hydroperoxide than the wt by disc inhibition assays at 42 degrees C, though minimum inhibitory concentration comparisons were inconclusive. Differences in genome-wide gene expression between wt and Delta luxS mutant with and without H(2)O(2) treatments were compared using DNA microarrays. The genes that showed differential expressions (wt/Delta luxS) include operons/pathways involved in AI-2 synthesis and S-adenosylmethionine (SAM) metabolism (metE, metF, and pfs), flagellar assembly/regulation, stress response (ahpC, tpx, and groES), ABC transporters/efflux systems, and two genes of unknown function located downstream of luxS (Cj1199 and Cj1200). The wt/Delta luxS expression ratios of ahpC (encoding alkyl hydroperoxide reductase) and tpx (encoding thiol peroxidase) were increased only with H(2)O(2) treatment, consistent with our finding that the Delta luxS mutant exhibits higher sensitivity to oxidative stress than wt. Our microarray results agreed with the Delta luxS mutant phenotypes, and suggested that LuxS plays a role in central metabolism involving SAM metabolism, but it is uncertain whether AI-2 functions as a true quorum-sensing signal in C. jejuni.
自诱导物-2(AI-2)是一种群体感应信号分子,可响应革兰氏阴性菌和革兰氏阳性菌中的细胞密度来控制多种细胞活动。AI-2的产生依赖于LuxS,它是AI-2生物合成途径中的最后一种酶。在本研究中,我们在空肠弯曲菌81-176菌株中构建了luxS无效突变体(ΔluxS),并表明它消除了AI-2的产生。与野生型(wt)相比,ΔluxS突变体在穆勒-欣顿(MH)肉汤中的倍增时间更长,并且在37℃的MH软琼脂上的群游能力降低,而在42℃时的生长速率或群游能力不受影响。通过42℃的纸片抑菌试验,ΔluxS突变体对过氧化氢(H₂O₂)和氢过氧化异丙苯也比wt更敏感,尽管最小抑菌浓度比较尚无定论。使用DNA微阵列比较了wt和ΔluxS突变体在有和没有H₂O₂处理情况下全基因组范围的基因表达差异。显示出差异表达(wt/ΔluxS)的基因包括参与AI-2合成和S-腺苷甲硫氨酸(SAM)代谢(metE、metF和pfs)的操纵子/途径、鞭毛组装/调节、应激反应(ahpC、tpx和groES)、ABC转运蛋白/外排系统,以及位于luxS下游的两个功能未知的基因(Cj1199和Cj1200)。ahpC(编码烷基过氧化氢还原酶)和tpx(编码硫醇过氧化物酶)的wt/ΔluxS表达比值仅在H₂O₂处理时增加,这与我们发现的ΔluxS突变体比wt对氧化应激表现出更高敏感性一致。我们的微阵列结果与ΔluxS突变体表型一致,并表明LuxS在涉及SAM代谢的中心代谢中起作用,但尚不确定AI-2在空肠弯曲菌中是否作为真正的群体感应信号发挥作用。