Quantitative proteomic analysis reveals that chemotaxis is involved in chlortetracycline resistance of Aeromonas hydrophila

Wanxin Li; Farman Ali; Qilan Cai; Zujie Yao; Lina Sun; Wenxiong Lin; Xiangmin Lin

doi:10.1016/j.jprot.2017.09.011

Quantitative proteomic analysis reveals that chemotaxis is involved in chlortetracycline resistance of Aeromonas hydrophila

J Proteomics. 2018 Feb 10:172:143-151. doi: 10.1016/j.jprot.2017.09.011. Epub 2017 Oct 3.

Authors

Wanxin Li¹, Farman Ali¹, Qilan Cai¹, Zujie Yao¹, Lina Sun¹, Wenxiong Lin¹, Xiangmin Lin²

Affiliations

¹ Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 35002, PR China; Key Laboratory of Crop Ecology and Molecular Physiology (Fujian Agriculture and Forestry University), Fujian Province University, Fuzhou 35002, PR China.
² Fujian Provincial Key Laboratory of Agroecological Processing and Safety Monitoring, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 35002, PR China; Key Laboratory of Crop Ecology and Molecular Physiology (Fujian Agriculture and Forestry University), Fujian Province University, Fuzhou 35002, PR China. Electronic address: xiangmin@fafu.edu.cn.

PMID: 28986269
DOI: 10.1016/j.jprot.2017.09.011

Abstract

In recent years, Aeromonas hydrophila, which has been classified as a food borne pathogen, has presented with increased levels of antibiotic resistance, with the mechanisms of this resistance being poorly understood. In this study, iTRAQ coupled mass spectrometry was employed to compare differentially expressed proteins in chlortetracycline (CTC) resistant A. hydrophila relative to a control strain. Result showed that a total of 234 differential proteins including 151 down-regulated and 83 up-regulated were identified in chlortetracycline resistance strain. Bioinformatics analysis showed that chemotaxis related proteins, such as CheA-2, CheR-3, CheW-2, EnvZ, PolA, FliS and FliG were down-regulated in addition to previously reported tricarboxylic acid cycle (TCA) related proteins also being down-regulated. A subset of identified differentially expressed proteins was then further validated via Western blotting. Exogenous metabolite combined with CTC further enhanced the bacterial susceptibilities to CTC in A. hydrophila. Furthermore, a bacterial survival capability assay showed that several chemotaxis related mutants, such as ΔcheR-3 and ΔAHA_0305, may affect the antimicrobial susceptibility of A. hydrophila. Overall, these findings contribute to a further understanding of the mechanism of CTC resistance in A. hydrophila and may contribute to the development of more effective future treatments.

Biological significance: A. hydrophila is a well-known fish pathogenic bacterium and has presented with increasing levels of antibiotic resistance, with the mechanisms of this resistance being poorly understood. Our current study compared the differentially expression proteins between chlortetracycline (CTC) resistant and control stains via an iTARQ-based quantitative proteomics method. Chemotaxis related proteins were down-regulated in CTC resistant strain but exogenous metabolite addition increased bacterial susceptibility in A.hydrophila. Significantly, chemotaxis related genes depletion affected antimicrobial susceptibilities of A.hydrophila indicating the role of chemotaxis process in antibiotics resistance.

Keywords: Aeromonas hydrophila; Antibiotic resistance; Chemotaxis; Quantitative proteomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aeromonas hydrophila / physiology*
Anti-Bacterial Agents
Bacterial Proteins / analysis
Bacterial Proteins / genetics
Chemotaxis / genetics*
Chlortetracycline / pharmacology*
Drug Resistance, Microbial / genetics*
Gene Expression Regulation, Bacterial
Gram-Negative Bacterial Infections
Proteomics / methods*

Substances

Anti-Bacterial Agents
Bacterial Proteins
Chlortetracycline