{"title":"Genome-Wide Screens Identify Genes Responsible for Intrinsic Boric Acid Resistance in Escherichia coli.","authors":"Bekir Çöl, Merve Sezer Kürkçü, Esra Di Bek","doi":"10.1007/s12011-024-04129-0","DOIUrl":null,"url":null,"abstract":"<p><p>Boric acid (BA) has antimicrobial properties and is used to combat bacterial infections, including Enterobacteria. However, the molecular mechanisms and cellular responses to BA are still unknown. This genomics study aims to provide new information on the genes and molecular mechanisms related to the antimicrobial effect of BA in Escherichia coli. The Keio collection of E. coli was used to screen 3985 single-gene knockout strains in order to identify mutant strains that were sensitive or hypersensitive to BA at certain concentrations. The mutant strains were exposed to different concentrations of BA ranging from 0 to 120 mM in LB media. Through genome-wide screens, 92 mutants were identified that were relatively sensitive to BA at least at one concentration tested. The related biological processes in the particular cellular system were listed. This study demonstrates that intrinsic BA resistance is the result of various mechanisms acting together. Additionally, we identified eighteen out of ninety-two mutant strains (Delta_aceF, aroK, cheZ, dinJ, galS, garP, glxK, nohA, talB, torR, trmU, trpR, yddE, yfeS, ygaV, ylaC, yoaC, yohN) that exhibited sensitivity using other methods. To increase sensitivity to BA, we constructed double and triple knockout mutants of the selected sensitive mutants. In certain instances, engineered double and triple mutants exhibited significantly amplified effects. Overall, our analysis of these findings offers further understanding of the mechanisms behind BA toxicity and intrinsic resistance in E. coli.</p>","PeriodicalId":8917,"journal":{"name":"Biological Trace Element Research","volume":" ","pages":"5771-5793"},"PeriodicalIF":3.4000,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11502571/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biological Trace Element Research","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s12011-024-04129-0","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/3/11 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Boric acid (BA) has antimicrobial properties and is used to combat bacterial infections, including Enterobacteria. However, the molecular mechanisms and cellular responses to BA are still unknown. This genomics study aims to provide new information on the genes and molecular mechanisms related to the antimicrobial effect of BA in Escherichia coli. The Keio collection of E. coli was used to screen 3985 single-gene knockout strains in order to identify mutant strains that were sensitive or hypersensitive to BA at certain concentrations. The mutant strains were exposed to different concentrations of BA ranging from 0 to 120 mM in LB media. Through genome-wide screens, 92 mutants were identified that were relatively sensitive to BA at least at one concentration tested. The related biological processes in the particular cellular system were listed. This study demonstrates that intrinsic BA resistance is the result of various mechanisms acting together. Additionally, we identified eighteen out of ninety-two mutant strains (Delta_aceF, aroK, cheZ, dinJ, galS, garP, glxK, nohA, talB, torR, trmU, trpR, yddE, yfeS, ygaV, ylaC, yoaC, yohN) that exhibited sensitivity using other methods. To increase sensitivity to BA, we constructed double and triple knockout mutants of the selected sensitive mutants. In certain instances, engineered double and triple mutants exhibited significantly amplified effects. Overall, our analysis of these findings offers further understanding of the mechanisms behind BA toxicity and intrinsic resistance in E. coli.
硼酸(BA)具有抗菌特性,可用于抗击包括肠杆菌在内的细菌感染。然而,人们对硼酸的分子机制和细胞反应仍一无所知。这项基因组学研究旨在为大肠杆菌中与 BA 的抗菌作用有关的基因和分子机制提供新的信息。研究人员利用庆应义塾大学收集的大肠杆菌筛选了 3985 株单个基因敲除菌株,以确定在特定浓度下对 BA 敏感或不敏感的突变菌株。突变菌株在 LB 培养基中暴露于 0 至 120 mM 不同浓度的 BA。通过全基因组筛选,确定了 92 个突变体,这些突变体至少在一个测试浓度下对 BA 相对敏感。列出了特定细胞系统中的相关生物过程。这项研究表明,BA 的内在抗性是各种机制共同作用的结果。此外,我们还从 92 个突变菌株(Delta_aceF、aroK、cheZ、dinJ、galS、garP、glxK、nohA、talB、torR、trmU、trpR、yddE、yfeS、ygaV、ylaC、yoaC、yohN)中发现了 18 个使用其他方法表现出敏感性的菌株。为了提高对 BA 的敏感性,我们构建了所选敏感突变体的双基因敲除突变体和三基因敲除突变体。在某些情况下,设计的双突变体和三突变体表现出明显的放大效应。总之,我们对这些发现的分析有助于进一步了解大肠杆菌中 BA 毒性和内在抗性背后的机制。
期刊介绍:
Biological Trace Element Research provides a much-needed central forum for the emergent, interdisciplinary field of research on the biological, environmental, and biomedical roles of trace elements. Rather than confine itself to biochemistry, the journal emphasizes the integrative aspects of trace metal research in all appropriate fields, publishing human and animal nutritional studies devoted to the fundamental chemistry and biochemistry at issue as well as to the elucidation of the relevant aspects of preventive medicine, epidemiology, clinical chemistry, agriculture, endocrinology, animal science, pharmacology, microbiology, toxicology, virology, marine biology, sensory physiology, developmental biology, and related fields.