{"title":"The dynamics of colicin E9 release from Escherichia coli in native conditions.","authors":"S. Bano, S. A. Tunio, C. Penfold, Richard James","doi":"10.1093/lambio/ovae042","DOIUrl":null,"url":null,"abstract":"Colicin (Col) plasmid contains colicin encoding genes arranged in an operon controlled by an SOS inducible promoter. Therefore, any external stresses to the host cell can induce the expression of the downstream genes in the Col operon including a lysis gene. The lysis protein is involved in the extracellular release of colicin through lysis of the producer cells which causes a decline in culture turbidity. However, it is not yet known that E. coli cells with ColE9-J plasmid hold the same level of cell death at the population level following a set of induced conditions. In this study, using a mitomycin C sensitivity assay along with a live dead staining method of detection, we showed that the native pColE9-J plasmid which unusually carries an extended Col operon (ColE9) containing two lysis genes, did not confer a rapid decline in the culture turbidity following induction with mitomycin C. Interestingly a subset of the cells suffered perturbation of their outer membrane which was not observed from single lysis mutant (∆celE or ∆celI) cells. This observed heterogeneity in the ColE9 release leading to differential outer membrane perforation may bring a competitive advantage to these cells in a mixed population.","PeriodicalId":2,"journal":{"name":"ACS Applied Bio Materials","volume":null,"pages":null},"PeriodicalIF":4.6000,"publicationDate":"2024-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Applied Bio Materials","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/lambio/ovae042","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MATERIALS SCIENCE, BIOMATERIALS","Score":null,"Total":0}
引用次数: 0
Abstract
Colicin (Col) plasmid contains colicin encoding genes arranged in an operon controlled by an SOS inducible promoter. Therefore, any external stresses to the host cell can induce the expression of the downstream genes in the Col operon including a lysis gene. The lysis protein is involved in the extracellular release of colicin through lysis of the producer cells which causes a decline in culture turbidity. However, it is not yet known that E. coli cells with ColE9-J plasmid hold the same level of cell death at the population level following a set of induced conditions. In this study, using a mitomycin C sensitivity assay along with a live dead staining method of detection, we showed that the native pColE9-J plasmid which unusually carries an extended Col operon (ColE9) containing two lysis genes, did not confer a rapid decline in the culture turbidity following induction with mitomycin C. Interestingly a subset of the cells suffered perturbation of their outer membrane which was not observed from single lysis mutant (∆celE or ∆celI) cells. This observed heterogeneity in the ColE9 release leading to differential outer membrane perforation may bring a competitive advantage to these cells in a mixed population.
可乐菌素(Col)质粒含有可乐菌素编码基因,这些基因排列在由 SOS 诱导型启动子控制的操作子中。因此,宿主细胞受到的任何外部压力都能诱导 Col 操作子中下游基因(包括裂解基因)的表达。裂解蛋白通过裂解生产者细胞参与细胞外秋水仙素的释放,从而导致培养液浊度下降。然而,目前还不清楚带有 ColE9-J 质粒的大肠杆菌细胞在一系列诱导条件下是否会在群体水平上保持相同的细胞死亡水平。在这项研究中,我们使用丝裂霉素 C 敏感性检测法和活死细胞染色检测法,结果表明,原生 pColE9-J 质粒异常地携带了一个包含两个裂解基因的扩展 Col 操作子(ColE9),在丝裂霉素 C 诱导后,培养液浊度并没有迅速下降。观察到的 ColE9 释放导致不同外膜穿孔的异质性可能会给混合群体中的这些细胞带来竞争优势。