Onufryk Christina, Crouch Marie-Laure, Fang Ferric C, Gross Carol A
Department of Biochemistry & Biophysics, University of California, San Francisco, California 94143, USA.
J Bacteriol. 2005 Jul;187(13):4552-61. doi: 10.1128/JB.187.13.4552-4561.2005.
In Escherichia coli, sigma(E) regulon functions are required for envelope homeostasis during stress and are essential for viability under all growth conditions. The E. coli genome encodes approximately 100 lipoproteins, and 6 of these are regulated by sigma(E). Phenotypes associated with deletion of each of these lipoproteins are the subject of this report. One lipoprotein, YfiO, is essential for cellular viability. However, overexpression of this protein is not sufficient to alleviate the requirement of sigma(E) for viability, suggesting that the sigma(E) regulon provides more than one essential function. The remaining five lipoproteins in the sigma(E) regulon are nonessential; cells are viable even when all five are removed simultaneously. Deletion of three nonessential lipoprotein genes (nlpB, yraP, ygfL) results in the exhibition of phenotypes that suggest they are important for maintenance of the integrity of the cell envelope. deltanlpB cells are selectively sensitive to rifampin; deltayraP cells are selectively sensitive to sodium dodecyl sulfate. Such selective sensitivity has not been previously reported. Both deltayraP and deltanlpB are synthetically lethal with surA::Cm, which encodes a periplasmic chaperone and PPIase, suggesting that NlpB and YraP play roles in a periplasmic folding pathway that functions in parallel with that of SurA. Finally, the deltayfgL mutant exhibits a broad range of envelope defects, including sensitivity to several membrane-impermeable agents, an altered outer membrane protein profile, synthetic lethality with both surA::Cm and deltafkpA::Cm strains, and sensitivity to a bactericidal permeability-increasing peptide. We suggest that this lipoprotein performs a very important but as-yet-unknown function in maintaining the integrity of the cell envelope.
在大肠杆菌中,σ(E) 调控子功能在应激期间对于包膜稳态是必需的,并且在所有生长条件下对于生存能力都是至关重要的。大肠杆菌基因组编码约100种脂蛋白,其中6种受σ(E) 调控。本报告的主题是与这些脂蛋白各自缺失相关的表型。一种脂蛋白YfiO对于细胞生存能力是必需的。然而,该蛋白的过表达不足以减轻σ(E) 对生存能力的需求,这表明σ(E) 调控子提供不止一种基本功能。σ(E) 调控子中的其余五种脂蛋白是非必需的;即使同时去除所有五种,细胞仍可存活。缺失三个非必需脂蛋白基因(nlpB、yraP、ygfL)会导致表现出一些表型,这表明它们对于维持细胞膜完整性很重要。nlpB缺失细胞对利福平具有选择性敏感性;yraP缺失细胞对十二烷基硫酸钠具有选择性敏感性。这种选择性敏感性以前尚未有报道。yraP缺失和nlpB缺失与编码周质伴侣和肽脯氨酰顺反异构酶的surA::Cm都是合成致死的,这表明NlpB和YraP在与SurA并行发挥作用的周质折叠途径中发挥作用。最后,ygfL缺失突变体表现出广泛的包膜缺陷,包括对几种膜不透性试剂敏感、外膜蛋白谱改变、与surA::Cm和fkpA::Cm缺失菌株都是合成致死的,以及对一种杀菌通透性增加肽敏感。我们认为这种脂蛋白在维持细胞膜完整性方面发挥着非常重要但尚未知的功能。