title: Viability, morphology, and proteome of Mycobacterium smegmatis MSMEG_0319 knockout strain.
abstract: Mycobacterium tuberculosis Rv0228, a membrane protein, is predicted as a drug target through computational methods. MSMEG_0319 (MS0319) in Mycobacterium smegmatis mc(2) 155 is the ortholog of Rv0228. To study the effect of MS0319 protein on M. smegmatis, an MS0319 gene knockout strain (ΔMS0319) was generated via a homologous recombination technique in this study. The results showed that the lack of MS0319 protein in mc(2) 155 cells led to the loss of viability at nonpermissive temperature. Scanning electron microscopy and transmission electron microscopy observations showed drastic changes in cellular shape especially cell wall disruption in ΔMS0319 cells. Proteomic analysis of ΔMS0319 cells through LC-MS/MS revealed that 462 proteins had changes in their expressions by lacking MS0319 protein. The M. tuberculosis orthologs of these 462 proteins were found through BLASTp search and functional clustering and metabolic pathway enrichment were performed on the orthologs. The results revealed that most of them were enzymes involved in metabolism of carbohydrates and amino acids, indicating that Rv0228 played an important role in cellular metabolism. All these results suggested Rv0228 as a potential target for development of antituberculosis drugs.
authors.affiliation: Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, P. R. China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Sha
authors.affiliation: Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, P. R. China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Shi
authors.affiliation: Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, P. R. China.
authors.forename: Liming
authors.initials: L
authors.lastname: Xu
authors.affiliation: Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, P. R. China.
authors.forename: Jiabin
authors.initials: J
authors.lastname: Wen
authors.affiliation: Department of Biotechnology, Dalian Medical University, Dalian, P. R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Xin
authors.affiliation: Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, P. R. China.
authors.forename: Yufang
authors.initials: Y
authors.lastname: Ma
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D016000:Cluster Analysis; D055808:Drug Discovery; D055786:Gene Knockout Techniques; D006358:Hot Temperature; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D050296:Microbial Viability / Q000235:genetics; D020102:Mycobacterium smegmatis / Q000235:genetics* / Q000502:physiology*; D020543:Proteome / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D020543:Proteome
keywords: Drug target; M. smegmatis; M. tuberculosis; MSMEG_0319; Membrane protein; Microbiology; Rv0228