Single cell analysis of Vibrio harveyi uncovers functional heterogeneity in response to quorum sensing signals.
Single cell analysis of Vibrio harveyi uncovers functional heterogeneity in response to quorum sensing signals.
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DOI：
10.1186/1471-2180-12-209
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发表时间：
2012-09-18
期刊：
影响因子：
4.2
通讯作者：
Jung K
中科院分区：
文献类型：
--
作者：
Anetzberger C;Schell U;Jung K
哈维氏弧菌及其近缘种是水产养殖的重要病原菌。一个复杂的群体感应级联涉及三个自诱导控制生物发光和几个基因编码的毒力因子。哈维氏弧菌种群的单细胞分析已经表明生物发光产生的细胞间异质性。本研究旨在分析单个细胞中各种自身诱导物依赖性基因的表达。在这里，我们使用携带启动子：：gfp融合的报告菌株来监测在单细胞水平上野生型缀合物中三个自诱导物调节基因的诱导/抑制。选择参与发病机制的两个基因-分别编码外切蛋白酶和III型分泌系统的组分的vhp和vscP以及luxC（lux操纵子中的第一个基因）进行分析。lux操纵子和外切蛋白酶基因被诱导，而vscP在高细胞密度下被抑制。作为对照，检查luxS和recA，其表达不依赖于自身诱导物。启动子：：gfp融合体在来自相同培养物的单个细胞中的反应从无诱导到高诱导。重要的是，在单细胞中同时分析两种自诱导物诱导的表型，生物发光（光检测）和胞外蛋白水解活性（启动子：：gfp融合的荧光），为哈维氏弧菌群体内的功能异质性提供了证据。自诱导物不仅是细胞密度的指示物，而且在群体内生理活动的协调中起着关键作用。
Vibrio harveyi and closely related species are important pathogens in aquaculture. A complex quorum sensing cascade involving three autoinducers controls bioluminescence and several genes encoding virulence factors. Single cell analysis of a V. harveyi population has already indicated intercellular heterogeneity in the production of bioluminescence. This study was undertaken to analyze the expression of various autoinducer-dependent genes in individual cells. Here we used reporter strains bearing promoter::gfp fusions to monitor the induction/repression of three autoinducer-regulated genes in wild type conjugates at the single cell level. Two genes involved in pathogenesis - vhp and vscP, which code for an exoprotease and a component of the type III secretion system, respectively, and luxC (the first gene in the lux operon) were chosen for analysis. The lux operon and the exoprotease gene are induced, while vscP is repressed at high cell density. As controls luxS and recA, whose expression is not dependent on autoinducers, were examined. The responses of the promoter::gfp fusions in individual cells from the same culture ranged from no to high induction. Importantly, simultaneous analysis of two autoinducer induced phenotypes, bioluminescence (light detection) and exoproteolytic activity (fluorescence of a promoter::gfp fusion), in single cells provided evidence for functional heterogeneity within a V. harveyi population. Autoinducers are not only an indicator for cell density, but play a pivotal role in the coordination of physiological activities within the population.
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影响因子：
2.5
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Austin, B;Pride, AC;Rhodie, GA
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