title: Proteomic Analysis of Drug-Resistant Mycobacteria: Co-Evolution of Copper and INH Resistance.
abstract: Tuberculosis, caused by the pathogen Mycobacterium tuberculosis, is a worldwide public health threat. Mycobacterium tuberculosis is capable of resisting various stresses in host cells, including high levels of ROS and copper ions. To better understand the resistance mechanisms of mycobacteria to copper, we generated a copper-resistant strain of Mycobacterium smegmatis, mc2155-Cu from the selection of copper sulfate treated-bacteria. The mc2155-Cu strain has a 5-fold higher resistance to copper sulfate and a 2-fold higher resistance to isoniazid (INH) than its parental strain mc2155, respectively. Quantitative proteomics was carried out to find differentially expressed proteins between mc2155 and mc2155-Cu. Among 345 differentially expressed proteins, copper-translocating P-type ATPase was up-regulated, while all other ABC transporters were down-regulated in mc2155-Cu, suggesting copper-translocating P-type ATPase plays a crucial role in copper resistance. Results also indicated that the down-regulation of metabolic enzymes and decreases in cellular NAD, FAD, mycothiol, and glutamine levels in mc2155-Cu were responsible for its slowing growth rate as compared to mc2155. Down-regulation of KatG2 expression in both protein and mRNA levels indicates the co-evolution of copper and INH resistance in copper resistance bacteria, and provides new evidence to understanding of the molecular mechanisms of survival of mycobacteria under stress conditions.
authors.affiliation: MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Chen
authors.affiliation: MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China.
authors.forename: Zhongyuan
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Beijing Chaoyang Hospital Affiliated to Capital Medical University, Beijing, China.
authors.forename: Qingtao
authors.initials: Q
authors.lastname: Wang
authors.affiliation: CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, CAS, Beijing, 100101, China.
authors.forename: Kaixia
authors.initials: K
authors.lastname: Mi
authors.affiliation: MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, China.
authors.forename: Haiteng
authors.initials: H
authors.lastname: Deng
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism*; D003300:Copper / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial; D006801:Humans; D007538:Isoniazid / Q000494:pharmacology*; D055432:Metabolomics / Q000379:methods; D009165:Mycobacterium Infections, Nontuberculous / Q000188:drug therapy / Q000378:metabolism* / Q000382:microbiology; D020102:Mycobacterium smegmatis / Q000187:drug effects / Q000502:physiology*; D040901:Proteomics / Q000379:methods*
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D003300:Copper; D007538:Isoniazid