Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
Mol Microbiol. 2021 Feb;115(2):191-207. doi: 10.1111/mmi.14609. Epub 2020 Oct 12.
Undecaprenyl phosphate (Und-P) is an essential lipid carrier that ferries cell wall intermediates across the cytoplasmic membrane in bacteria. Und-P is generated by dephosphorylating undecaprenyl pyrophosphate (Und-PP). In Escherichia coli, BacA, PgpB, YbjG, and LpxT dephosphorylate Und-PP and are conditionally essential. To identify vulnerabilities that arise when Und-P metabolism is defective, we developed a genetic screen for synthetic interactions which, in combination with ΔybjG ΔlpxT ΔbacA, are lethal or reduce fitness. The screen uncovered novel connections to cell division, DNA replication/repair, signal transduction, and glutathione metabolism. Further analysis revealed several new morphogenes; loss of one of these, qseC, caused cells to enlarge and lyse. QseC is the sensor kinase component of the QseBC two-component system. Loss of QseC causes overactivation of the QseB response regulator by PmrB cross-phosphorylation. Here, we show that deleting qseB completely reverses the shape defect of ΔqseC cells, as does overexpressing rprA (a small RNA). Surprisingly, deleting pmrB only partially suppressed qseC-related shape defects. Thus, QseB is activated by multiple factors in QseC's absence and prior functions ascribed to QseBC may originate from cell wall defects. Altogether, our findings provide a framework for identifying new determinants of cell integrity that could be targeted in future therapies.
十一烯基磷酸酯(Und-P)是一种必需的脂质载体,可在细菌中通过细胞质膜运输细胞壁中间体。Und-P 是通过去磷酸化十一烯基焦磷酸酯(Und-PP)产生的。在大肠杆菌中,BacA、PgpB、YbjG 和 LpxT 去磷酸化 Und-PP,是条件必需的。为了确定 Und-P 代谢缺陷时出现的脆弱性,我们开发了一种用于合成相互作用的遗传筛选,该筛选与ΔybjGΔlpxTΔbacA 结合使用是致命的或降低适应性。该筛选揭示了与细胞分裂、DNA 复制/修复、信号转导和谷胱甘肽代谢的新联系。进一步的分析显示了几个新的形态发生基因;其中一个基因(qseC)的缺失导致细胞增大和裂解。QseC 是 QseBC 双组分系统的传感器激酶成分。QseC 的缺失导致 PmrB 交叉磷酸化过度激活 QseB 应答调节子。在这里,我们表明,完全删除 qseB 可完全逆转ΔqseC 细胞的形状缺陷,rprA(一种小 RNA)的过表达也是如此。令人惊讶的是,删除 pmrB 只能部分抑制 qseC 相关的形状缺陷。因此,在 QseC 缺失的情况下,QseB 被多种因素激活,而先前归因于 QseBC 的功能可能源于细胞壁缺陷。总之,我们的研究结果为确定新的细胞完整性决定因素提供了一个框架,这些决定因素可能成为未来治疗的靶点。