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LuxR- and Acyl-Homoserine-Lactone-Controlled Non-lux Genes Define a Quorum-Sensing Regulon in Vibrio fischeri

机译:LuxR和酰基高苏氨酸-内酯控制的非lux基因在费氏弧菌中定义了群体感应调节子。

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摘要

The luminescence (lux) operon (luxICDABEG) of the symbiotic bacterium Vibrio fischeri is regulated by the transcriptional activator LuxR and two acyl-homoserine lactone (acyl-HSL) autoinducers (the luxI-dependent 3-oxo-hexanoyl-HSL [3-oxo-C6-HSL] and the ainS-dependent octanoyl-HSL [C8-HSL]) in a population density-responsive manner called quorum sensing. To identify quorum-sensing-regulated (QSR) proteins different from those encoded by lux genes, we examined the protein patterns of V. fischeri quorum-sensing mutants defective in luxI, ainS, and luxR by two-dimensional polyacrylamide gel electrophoresis. Five non-Lux QSR proteins, QsrP, RibB, AcfA, QsrV, and QSR 7, were identified; their production occurred preferentially at high population density, required both LuxR and 3-oxo-C6-HSL, and was inhibited by C8-HSL at low population density. The genes encoding two of the QSR proteins were characterized: qsrP directs cells to synthesize an apparently novel periplasmic protein, and ribB is a homolog of the Escherichia coli gene for 3,4-dihydroxy-2-butanone 4-phosphate synthase, a key enzyme for riboflavin synthesis. The qsrP and ribB promoter regions each contained a sequence similar to the lux operon lux box, a 20-bp region of dyad symmetry necessary for LuxR/3-oxo-C6-HSL-dependent activation of lux operon transcription. V. fischeri qsrP and ribB mutants exhibited no distinct phenotype in culture. However, a qsrP mutant, in competition with its parent strain, was less successful in colonizing Euprymna scolopes, the symbiotic host of V. fischeri. The newly identified QSR genes, together with the lux operon, define a LuxR/acyl-HSL-responsive quorum-sensing regulon in V. fischeri.
机译:共生细菌费氏弧菌的发光(lux)操纵子(luxICDABEG)受转录激活因子LuxR和两个酰基-高丝氨酸内酯(酰基-HSL)自动诱导因子(依赖于luxI的3-氧代-己基-HSL [3-oxo -C6-HSL]和依赖ainS的辛酰基-HSL [C8-HSL]),以群体密度响应的方式称为群体感应。为了鉴定不同于lux基因编码的群体感应调节(QSR)蛋白,我们通过二维聚丙烯酰胺凝胶电泳检查了luxI,ainS和luxR缺陷的费氏弧菌群体感应突变体的蛋白质模式。确定了五个非Lux QSR蛋白,即QsrP,RibB,AcfA,QsrV和QSR 7。它们的产生优先在高种群密度下发生,需要LuxR和3-oxo-C6-HSL,并且在低种群密度下被C8-HSL抑制。表征了两种QSR蛋白的基因:qsrP指导细胞合成表面上新颖的周质蛋白,ribB是3,4-二羟基-2-丁酮4-磷酸合酶(一种关键酶)的大肠杆菌基因的同源物。用于核黄素合成。每个qsrP和ribB启动子区域都包含一个与lux操纵子lux盒类似的序列,这是LuxR / 3-oxo-C6-HSL依赖的lux操纵子转录激活所必需的20 bp dyad对称区域。费氏弧菌qsrP和ribB突变体在培养物中没有表现出明显的表型。然而,一个qsrP突变体与其亲本竞争,在定植费氏弧菌的共生宿主Euprymna scolopes方面不太成功。新近鉴定的QSR基因与勒克斯操纵子一起,在费氏弧菌中定义了LuxR /酰基-HSL反应性群体感应调节子。

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