Microbiology Department, The University of Iowa, Iowa City, IA 52242, USA.
J Bacteriol. 2011 Aug;193(16):4224-37. doi: 10.1128/JB.00432-11. Epub 2011 Jun 24.
The quorum regulatory cascade is poorly characterized in Vibrio parahaemolyticus, in part because swarming and virulence factors--the hallmarks of the organism--are repressed by this scheme of gene control, and quorum sensing seems to be silenced in many isolates. In these studies, we examine a swarming-proficient, virulent strain and identify an altered-function allele of the quorum regulator luxO that is demonstrated to produce a constitutively active mimic of LuxO∼P. We find that LuxO* affects the expression of three small regulatory RNAs (Qrrs) and the activity of a translational fusion in opaR, the output regulator. Tests for epistasis showed that luxO* is dominant over luxO and that opaR is dominant over luxO. Thus, information flow through the central elements of the V. parahaemolyticus quorum pathway is proven for the first time. Quorum-sensing output was explored using microarray profiling: the OpaR regulon encompasses ∼5.2% of the genome. OpaR represses the surface-sensing and type III secretion system 1 (T3SS1) regulons. One novel discovery is that OpaR strongly and oppositely regulates two type VI secretion systems (T6SS). New functional consequences of OpaR control were demonstrated: OpaR increases the cellular cyclic di-GMP (c-di-GMP) level, positively controls chitin-induced DNA competency, and profoundly blocks cytotoxicity toward host cells. In expanding the previously known quorum effects beyond the induction of the capsule and the repression of swarming to elucidate the global scope of genes in the OpaR regulon, this study yields many clues to distinguishing traits of this Vibrio species; it underscores the profoundly divergent survival strategies of the quorum On/Off phase variants.
群体感应调控级联在副溶血性弧菌中描述得很差,部分原因是群集和毒力因子——该生物体的标志——受到这种基因调控方案的抑制,并且群体感应似乎在许多分离株中被沉默。在这些研究中,我们研究了一个群集能力强、毒力强的菌株,并鉴定了群体感应调节剂 luxO 的一个改变功能的等位基因,该等位基因被证明产生了一个组成型激活的 LuxO∼P 模拟物。我们发现 LuxO* 影响三个小调节 RNA(Qrrs)的表达和 opaR 中的翻译融合的活性,opaR 是输出调节剂。上位性测试表明,luxO* 在 luxO 上是显性的,opaR 在 luxO 上是显性的。因此,首次证明了 V. parahaemolyticus 群体感应途径的中心元件的信息流。使用微阵列分析探索了群体感应输出:OpaR 调控子涵盖基因组的约 5.2%。OpaR 抑制表面感应和 III 型分泌系统 1(T3SS1)调控子。一个新的发现是 OpaR 强烈且相反地调控两个 VI 型分泌系统(T6SS)。OpaR 控制的新功能后果得到了证明:OpaR 增加细胞环二鸟苷酸(c-di-GMP)水平,正向控制几丁质诱导的 DNA 能力,并强烈阻止对宿主细胞的细胞毒性。在扩大先前已知的群体感应效应超出了对荚膜的诱导和群集的抑制,以阐明 OpaR 调控子中基因的全局范围,这项研究为区分该弧菌物种的特征提供了许多线索;它强调了群体感应 On/Off 相位变体的生存策略的深刻分歧。