Identification of Novel Genes Involved in Escherichia coli Persistence to Tosufloxacin

Abstract

Persisters are metabolically quiescent phenotypic variants of the wild type that are tolerant to cidal antibiotics, and the mechanisms of persister formation and survival are complex and not completely understood. To identify genes involved in persistence to tosufloxacin, which has higher activity against persisters than most other quinolones, we screened the E. coli KEIO mutant library using a different condition from most persister mutant screens (6 h) with a longer exposure of 18 h with tosufloxacin. We identified 18 mutants (

Rethinking Dormancy: Antibiotic Persisters are Metabolically Active, Non-Growing Cells

“…This observation demonstrated that the commonly upregulated genes follow consistency with different types of antibiotics; Amp targets cell wall formation 21 , while Cip targets DNA synthesis 41 .Our goal was to determine how these genes were related to persister survival. We build off what we knew from previous persister and transcriptomic analysis studies 24,[42][43][44][45][46][47][48] . Previous studies identified several persister-related genes including oxyR, dnaK, sucB, relA, rpoS, clpB, mqsR, and recA, which were found to be related to prominent persister formation with multiple antibiotics treatment 42 .…”

“…1. The fact that persister-related genes are constantly identified, and if one mechanism is reasoned for persister formation by one research group, another's group disclaimed that mechanism [42][43][44][45][46]71 . This suggests not one gene is responsible for persister formation.…”

“…In this manner, the condition that the cell entered the ground state determines the type of persister that will form. We make this hypothesis based on the fact that persister-related genes are constantly identified, and if one mechanism is reasoned for persister formation by one research group, another's group disclaimed that mechanism [42][43][44][45][46]68 (suggesting not one gene is responsible for persister formation). The minimal genome contains mainly essential genes (less than 500 genes) and still form persisters (other microbes contain genes that have been to be persister-related, but the minimal genome lacks them) 69 , slowed growth either externally 64 or internally 2,70 appears to increase persister levels (showing that both internal and external stress alters persister levels).…”

Enolase Is Implicated in the Emergence of Gonococcal Tolerance to Ceftriaxone

“…Mutations in tatC and edd genes were identified at timepoint 7 in lineage 1. Mutations in tatC and other genes, gltI, hlpA , ruvC , ddlB and ydfI, were found to result in tolerance to tosufloxacin in E. coli [ 27 ]. TatC, the primary substrate receptor, is part of the twin-arginine translocation (Tat) system needed to transport folded proteins across biological membranes [ 28 ].…”

Glutamate Transporters GltS, GltP and GltI Are Involved in Escherichia coli Tolerance In Vitro and Pathogenicity in Mouse Urinary Tract Infections

“…Compared with other quinolones, tosufloxacin had higher activity against E. coli tolerant persister cells [ 25 ]. In our previous work to identify genes involved in E. coli persistence, we found that mutation in the gltI encoding glutamate/aspartate transport protein showed increased susceptibility to tosufloxacin [ 26 ], suggesting that glutamate transporters may be involved in bacterial tolerance. In this study, to evaluate the role of glutamate transporter GltS, as well as two other glutamate transporters, GltP and GltI, in E. coli tolerance and pathogenicity, we constructed their deletion mutants in both E. coli K12 strain BW25113 and uropathogenic E. coli UTI89, followed by analysis of their tolerance to various antibiotics and stressors in vitro and their pathogenicity in human bladder epithelial cells and mouse urinary tract infections.…”