Derzelle Sylviane, Ngo Saravuth, Turlin Évelyne, Duchaud Éric, Namane Abdelkader, Kunst Frank, Danchin Antoine, Bertin Philippe, Charles Jean-François
Unité de Génétique des Génomes Bactériens, Département de Structure et Dynamique des Génomes, Institut Pasteur, 75724 Paris Cedex 15, France.
Laboratoire de Génomique des Micro-organismes Pathogènes, Département de Structure et Dynamique des Génomes, Institut Pasteur, 75724 Paris Cedex 15, France.
Microbiology (Reading). 2004 Apr;150(Pt 4):897-910. doi: 10.1099/mic.0.26563-0.
Photorhabdus luminescens is an insect-pathogenic bacterium that forms a symbiosis with specific entomopathogenic nematodes. In this bacterium, a symbiosis-'deficient' phenotypic variant (known as the secondary variant or form II) arises at a low frequency during prolonged incubation. A knock-out mutant was generated of the regulator of a newly identified two-component regulatory system, designated AstR-AstS. Interestingly, this mutation altered the timing of phenotypic switching. Variant cells arose in the mutant strain several days before they did in the wild-type population, suggesting that AstRS is directly or indirectly involved in the genetic mechanism underlying variant cell formation. This mutation also affected motility and antibiotic synthesis. To identify AstRS-regulated genes, a comparative analysis using two-dimensional gel electrophoresis was performed. Seventeen proteins with modified synthesis in stationary phase were identified by mass spectrometry and shown to be involved in electron-transport systems, energy metabolism, iron acquisition and stress responses. The results imply that AstRS is involved in the adaptation of cells to the stationary phase, whilst negatively affecting the competitive advantage of form I cells. The link between AstRS-dependent stationary-phase adaptation and phenotypic variation is discussed.
发光光杆状菌是一种昆虫致病细菌,它与特定的昆虫病原线虫形成共生关系。在这种细菌中,长时间培养时会以低频率出现一种共生缺陷型表型变体(称为次生变体或II型)。针对新鉴定的双组分调节系统AstR-AstS的调节因子构建了一个基因敲除突变体。有趣的是,这种突变改变了表型转换的时间。突变菌株中变体细胞的出现比野生型群体中早几天,这表明AstRS直接或间接参与了变体细胞形成的遗传机制。这种突变还影响了运动性和抗生素合成。为了鉴定受AstRS调节的基因,进行了二维凝胶电泳的比较分析。通过质谱鉴定了17种在稳定期合成发生改变的蛋白质,这些蛋白质参与电子传递系统、能量代谢、铁摄取和应激反应。结果表明,AstRS参与细胞对稳定期的适应,同时对I型细胞的竞争优势产生负面影响。本文还讨论了AstRS依赖性稳定期适应与表型变异之间的联系。