Competitive Growth Enhances Conditional Growth Mutant Sensitivity to Antibiotics and Exposes a Two-Component System as an Emerging Antibacterial Target in Burkholderia cenocepacia

Author:

Gislason April S.1,Choy Matthew1,Bloodworth Ruhi A. M.1,Qu Wubin2,Stietz Maria S.1,Li Xuan3,Zhang Chenggang2,Cardona Silvia T.14ORCID

Affiliation:

1. Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada

2. Beijing Institute of Radiation Medicine, State Key Laboratory of Proteomics, Cognitive and Mental Health Research Center, Beijing, China

3. Department of Mathematics and Statistics, University of Minnesota Duluth, Duluth, Minnesota, USA

4. Department of Medical Microbiology and Infectious Disease, University of Manitoba, Winnipeg, Canada

Abstract

ABSTRACT Chemogenetic approaches to profile an antibiotic mode of action are based on detecting differential sensitivities of engineered bacterial strains in which the antibacterial target (usually encoded by an essential gene) or an associated process is regulated. We previously developed an essential-gene knockdown mutant library in the multidrug-resistant Burkholderia cenocepacia by transposon delivery of a rhamnose-inducible promoter. In this work, we used Illumina sequencing of multiplex-PCR-amplified transposon junctions to track individual mutants during pooled growth in the presence of antibiotics. We found that competition from nontarget mutants magnified the hypersensitivity of a clone underexpressing gyrB to novobiocin by 8-fold compared with hypersensitivity measured during clonal growth. Additional profiling of various antibiotics against a pilot library representing most categories of essential genes revealed a two-component system with unknown function, which, upon depletion of the response regulator, sensitized B. cenocepacia to novobiocin, ciprofloxacin, tetracycline, chloramphenicol, kanamycin, meropenem, and carbonyl cyanide 3-chlorophenylhydrazone, but not to colistin, hydrogen peroxide, and dimethyl sulfoxide. We named the gene cluster esaSR for e nhanced s ensitivity to a ntibiotics s ensor and r esponse regulator. Mutational analysis and efflux activity assays revealed that while esaS is not essential and is involved in antibiotic-induced efflux, esaR is an essential gene and regulates efflux independently of antibiotic-mediated induction. Furthermore, microscopic analysis of cells stained with propidium iodide provided evidence that depletion of EsaR has a profound effect on the integrity of cell membranes. In summary, we unraveled a previously uncharacterized two-component system that can be targeted to reduce antibiotic resistance in B. cenocepacia .

Funder

Cystic Fibrosis Canada

Publisher

American Society for Microbiology

Subject

Infectious Diseases,Pharmacology (medical),Pharmacology

Reference84 articles.

1. Tackling antibiotic resistance

2. Bad Bugs, No Drugs: No ESKAPE! An Update from the Infectious Diseases Society of America

3. The multifarious, multireplicon Burkholderia cepacia complex

4. Burns JL . 2007. Antibiotic resistance of Burkholderia spp., p 81–91. InCoenyeT VandammeP (ed), Burkholderia: molecular microbiology and genomics. Horizon Bioscience, Norfolk, VA.

5. Response of Pseudomonas cepacia to beta-Lactam antibiotics: utilization of penicillin G as the carbon source

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3