Stutzmann Sandrine, Blokesch Melanie
Laboratory of Molecular Microbiology, Global Health Institute, School of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
mSphere. 2016 May 25;1(3). doi: 10.1128/mSphere.00098-16. eCollection 2016 May-Jun.
Vibrio cholerae, the causative agent of cholera, is a model organism for studying virulence regulation, biofilm formation, horizontal gene transfer, and the cell-to-cell communication known as quorum sensing (QS). As in any research field, discrepancies between data from diverse laboratories are sometimes observed for V. cholerae. Such discrepancies are often caused by the use of diverse patient or environmental isolates. In this study, we investigated the inability of a few laboratories to reproduce high levels of natural transformation, a mode of horizontal gene transfer that is specifically induced on chitinous surfaces. This irreproducibility was mostly related to one specific isolate of V. cholerae: the O1 El Tor C6706 strain. C6706 was previously described as QS proficient, an important prerequisite for the induction of natural competence for transformation. To elucidate the underlying problem, we collected seven isolates of the same C6706 strain from different research laboratories in North America and Europe and compared their phenotypes. Importantly, we observed a split response with respect to QS-related gene expression, including chitin-induced natural competence and type VI secretion (T6S). While approximately half of the strains behaved as reported for several other O1 El Tor pandemic isolates that are commonly studied in the laboratory, the other half were significantly impaired in QS-related expression patterns. This impairment was caused by a mutation in a QS-related gene (luxO). We conclude that the circulation of such QS-impaired wild-type strains is responsible for masking several important phenotypes of V. cholerae, including natural competence for transformation and T6S. IMPORTANCE Phenotypic diversity between laboratory-domesticated bacterial strains is a common problem and often results in the failed reproduction of published data. However, researchers rarely compare such strains to elucidate the underlying mutation(s). In this study, we tested one of the best-studied V. cholerae isolates, O1 El Tor strain C6706 (a patient isolate from Peru), with respect to two main phenotypes: natural competence for transformation and type VI secretion. We recently demonstrated that the two phenotypes are coregulated and specifically induced upon the growth of pandemic V. cholerae O1 El Tor strains on chitinous surfaces. We provide evidence that of seven C6706 strains collected from different laboratories, four were impaired in the tested phenotypes due to a mutation in a QS gene. Collectively, our data indicate that the circulation of such a mutated wild-type strain of C6706 might have had important consequences for QS-related data.
霍乱弧菌是霍乱的病原体,是研究毒力调控、生物膜形成、水平基因转移以及被称为群体感应(QS)的细胞间通讯的模式生物。与任何研究领域一样,霍乱弧菌不同实验室的数据有时会出现差异。这种差异通常是由于使用了不同的患者或环境分离株。在本研究中,我们调查了一些实验室无法重现高水平自然转化的情况,自然转化是一种在几丁质表面特异性诱导的水平基因转移模式。这种不可重复性主要与一种特定的霍乱弧菌分离株有关:O1 埃尔托 C6706 菌株。C6706 先前被描述为群体感应功能正常,这是诱导转化自然感受态的重要前提条件。为了阐明潜在问题,我们从北美和欧洲的不同研究实验室收集了同一 C6706 菌株的七个分离株,并比较了它们的表型。重要的是,我们观察到在与群体感应相关的基因表达方面出现了分歧反应,包括几丁质诱导的自然感受态和 VI 型分泌(T6S)。虽然大约一半的菌株表现与实验室中通常研究的其他几种 O1 埃尔托大流行分离株的报道一致,但另一半在与群体感应相关的表达模式上明显受损。这种损伤是由一个与群体感应相关的基因(luxO)发生突变引起的。我们得出结论,这种群体感应受损的野生型菌株的传播导致掩盖了霍乱弧菌的几种重要表型,包括转化自然感受态和 T6S。重要性实验室驯化的细菌菌株之间的表型多样性是一个常见问题,常常导致已发表数据无法重现。然而,研究人员很少比较此类菌株以阐明潜在的突变。在本研究中,我们针对两种主要表型测试了研究得最深入的霍乱弧菌分离株之一,O1 埃尔托菌株 C6706(一株来自秘鲁的患者分离株):转化自然感受态和 VI 型分泌。我们最近证明这两种表型是共同调控的,并且在大流行的霍乱弧菌 O1 埃尔托菌株在几丁质表面生长时特异性诱导。我们提供的证据表明,从不同实验室收集的七个 C6706 菌株中,有四个由于群体感应基因发生突变而在测试表型上受损。总体而言,我们的数据表明这种 C6706 突变野生型菌株的传播可能对与群体感应相关的数据产生了重要影响。