Small RNAs and Small Proteins Involved in Resistance to Cell Envelope Stress and Acid Shock in Escherichia coli: Analysis of a Bar-Coded Mutant Collection
Small RNAs and Small Proteins Involved in Resistance to Cell Envelope Stress and Acid Shock in Escherichia coli: Analysis of a Bar-Coded Mutant Collection
复制标题
DOI：
10.1128/jb.00873-09
复制
发表时间：
2010-01-01
影响因子：
3.2
通讯作者：
Storz, Gisela
中科院分区：
文献类型：
--
作者：
Hobbs, Errett C.;Astarita, Jillian L.;Storz, Gisela
大肠杆菌基因组编码了80多种小调控rna （sRNAs）和60种16至50个氨基酸的蛋白质（小蛋白质）。绝大多数相应的基因都没有已知的功能。我们筛选了125个DNA条形码突变体，以鉴定与sRNAs和小蛋白编码基因缺失相关的新型细胞包膜应激和急性酸休克表型。9个缺失突变体（ssrA、micA、ybaM、ryeF、yqcG、sroH、ybhT、yobF和glmY）对细胞包膜胁迫敏感，2个具有抗性（rybB和blr）。编码四种小蛋白（yqgB、mgrB、yobF和yceO）的基因缺失突变体对急性酸胁迫敏感。我们在与野生型lacZ突变细胞的一对一竞争分析中证实了这些表型。对SsrA表型的更详细的研究表明，核糖体的释放是抵抗细胞包膜应激的关键。我们生成的条形码删除集合可以筛选对几乎任何应力条件的敏感性或抗性。
More than 80 small regulatory RNAs (sRNAs) and 60 proteins of 16 to 50 amino acids (small proteins) are encoded in the Escherichia coli genome. The vast majority of the corresponding genes have no known function. We screened 125 DNA bar-coded mutants to identify novel cell envelope stress and acute acid shock phenotypes associated with deletions of genes coding for sRNAs and small proteins. Nine deletion mutants (ssrA, micA, ybaM, ryeF, yqcG, sroH, ybhT, yobF, and glmY) were sensitive to cell envelope stress and two were resistant (rybB and blr). Deletion mutants of genes coding for four small proteins (yqgB, mgrB, yobF, and yceO) were sensitive to acute acid stress. We confirmed each of these phenotypes in one-on-one competition assays against otherwise-wild-type lacZ mutant cells. A more detailed investigation of the SsrA phenotype suggests that ribosome release is critical for resistance to cell envelope stress. The bar-coded deletion collection we generated can be screened for sensitivity or resistance to virtually any stress condition.