title: The Wag31 protein interacts with AccA3 and coordinates cell wall lipid permeability and lipophilic drug resistance in Mycobacterium smegmatis.
abstract: Mycobacterium tuberculosis, especially drug resistant tuberculosis, is a serious threat to global human health. Compared with other bacterial pathogens, M. tuberculosis gains stronger natural drug resistance from its unusually lipid-rich cell wall. As a DivIVA homolog, Wag31 has been demonstrated to be closely involved in peptidoglycan synthesis, cell growth and cell division. Previous research rarely investigated the role of Wag31 in drug resistance. In this study, we found Wag31 knock-down in Mycobacterium smegmatis resulted in a co-decrease of the resistance to four lipophilic drugs (rifampicin, novobiocin, erythromycin and clofazimine) and an increase in the cell permeability to lipophilic molecules. Six proteins (AccA3, AccD4 and AccD5, Fas, InhA and MmpL3) that are involved in fatty acid and mycolic acid synthesis were identified in the Wag31 interactome through Co-Immunoprecipitation. The Wag31-AccA3 interaction was confirmed by the pull-down assay. AccA3 overexpression resulted in a decrease in lipid permeability and an increase in the resistance of rifampicin and novobiocin. It confirmed the close relationship of lipophilic drug resistance, lipid permeability and the Wag31-AccA3 interaction. These results demonstrated that Wag31 maintained the resistance to lipophilic drugs and that Wag31 could play a role in controlling the lipid permeability of the cell wall through the Wag31-AccA3 interaction.
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, PR China. Electronic address: wenxixu10@fudan.edu.cn.
authors.forename: Wen-xi
authors.initials: WX
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, PR China. Electronic address: zhanglu407@fudan.edu.cn.
authors.forename: Lu
authors.initials: L
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, PR China. Electronic address: 349623669@qq.com.
authors.forename: Jun-tao
authors.initials: JT
authors.lastname: Mai
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, PR China. Electronic address: 08300300048@fudan.edu.cn.
authors.forename: Ru-chao
authors.initials: RC
authors.lastname: Peng
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, PR China. Electronic address: 072023005@fudan.edu.cn.
authors.forename: En-zhuo
authors.initials: EZ
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, PR China. Electronic address: Chao.peng@merial.com.
authors.forename: Chao
authors.initials: C
authors.lastname: Peng
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai, PR China. Electronic address: hhwang@fudan.edu.cn.
authors.forename: Hong-hai
authors.initials: HH
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002463:Cell Membrane Permeability; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D055785:Gene Knockdown Techniques; D005798:Genes, Bacterial; D006801:Humans; D008563:Membrane Lipids / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D020102:Mycobacterium smegmatis / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000378:metabolism / Q000472:pathogenicity; D009675:Novobiocin / Q000494:pharmacology; D012293:Rifampin / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008563:Membrane Lipids; D009675:Novobiocin; D012293:Rifampin
keywords: ACCase; Ag84; Coiled-coil