Transcriptional regulation of opaR, qrr2-4 and aphA by the master quorum-sensing regulator OpaR in Vibrio parahaemolyticus.
Transcriptional regulation of opaR, qrr2-4 and aphA by the master quorum-sensing regulator OpaR in Vibrio parahaemolyticus.
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DOI：
10.1371/journal.pone.0034622
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Zhou D
中科院分区：
文献类型：
--
作者：
Zhang Y;Qiu Y;Tan Y;Guo Z;Yang R;Zhou D
副溶血性弧菌是通过粪-口途径引起感染性腹泻和肠胃炎的主要原因。哈维氏弧菌是鱼类和无脊椎动物的病原体，并已被用作群体感应（QS）研究的模型。LuxR是哈维氏弧菌的主QS调节因子（MQSR），并且其自身基因qrr 2 -4和aphA的LuxR依赖性表达已经在哈维氏弧菌中建立。副溶血性弧菌MQSR OpaR对靶基因的分子调控仍知之甚少。生物信息学分析表明，副溶血性弧菌OpaR、哈维氏弧菌LuxR、创伤弧菌SmcR和溶藻弧菌ValR是极其保守的，并且这四种MQSR似乎识别相同的保守顺式作用信号，这由在其靶启动子内表现为位置频率矩阵和20 bp盒的共有构建体表示。在副溶血性弧菌opaR、qrr 2 -4和aphA基因的上游DNA区域发现了MQSR盒样序列，引物延伸实验、凝胶迁移率变动实验和DNA酶I足迹分析等实验进一步证实了OpaR对这些靶基因的直接转录调控作用。翻译和转录开始，核心启动子元件的σ因子识别，Shine-Dalgarno序列的核糖体识别，和OpaR结合位点的OpaR的五个靶基因，这给出了OpaR依赖性启动子的结构图。进一步的计算机启动子分析表明，上述调控电路共享的其他几个密切相关的弧菌，但略有例外。本研究对OpaR在副溶血弧菌中对五个靶基因opaR、qrr 2 -4和ahpA的直接转录调控进行了全面的计算和表征。这些特征性的调节回路在哈维氏弧菌和副溶血性弧菌中是保守的。
Vibrio parahaemolyticus is a leading cause of infectious diarrhea and enterogastritis via the fecal-oral route. V. harveyi is a pathogen of fishes and invertebrates, and has been used as a model for quorum sensing (QS) studies. LuxR is the master QS regulator (MQSR) of V. harveyi, and LuxR-dependent expression of its own gene, qrr2–4 and aphA have been established in V. harveyi. Molecular regulation of target genes by the V. parahaemolyticus MQSR OpaR is still poorly understood. The bioinformatics analysis indicated that V. parahaemolyticus OpaR, V. harveyi LuxR, V. vulnificu SmcR, and V. alginolyticus ValR were extremely conserved, and that these four MQSRs appeared to recognize the same conserved cis-acting signals, which was represented by the consensus constructs manifesting as a position frequency matrix and as a 20 bp box, within their target promoters. The MQSR box-like sequences were found within the upstream DNA regions of opaR, qrr2–4 and aphA in V. parahaemolyticus, and the direct transcriptional regulation of these target genes by OpaR were further confirmed by multiple biochemical experiments including primer extension assay, gel mobility shift assay, and DNase I footprinting analysis. Translation and transcription starts, core promoter elements for sigma factor recognition, Shine-Dalgarno sequences for ribosome recognition, and OpaR-binding sites were determined for the five target genes of OpaR, which gave a structural map of the OpaR-dependent promoters. Further computational promoter analysis indicated the above regulatory circuits were shared by several other closely related Vibrios but with slight exceptions. This study gave a comprehensive computational and characterization of the direct transcriptional regulation of five target genes, opaR, qrr2–4 and ahpA, by OpaR in V. parahaemolyticus. These characterized regulatory circuits were conserved in V. harveyi and V. parahaemolyticus.
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作者：
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