title: Decreased intracellular survival of an fkpA mutant of Salmonella typhimurium Copenhagen.
abstract: The fkpA gene of Salmonella typhimurium encodes a protein similar to the macrophage infectivity potentiator (Mip) proteins of Legionella pneumophila and Chlamydia trachomatis. Because Mip proteins enhance the ability of these intracellular pathogens to survive within macrophages and epithelial cells, we tested whether the product of the fkpA gene would have the same effect on the intracellular growth of a virulent strain of S. typhimurium. By a series of P22 transductions, the fkpA gene of S. typhimurium Copenhagen was replaced with the inactive fkpA1::omega-Cm gene from Escherichia coli, creating the mutant S. typhimurium KY32H1. The Copenhagen and KY32H1 strains were equally able to enter Caco-2 cells (an epithelial cell line) and J774.A1 cells (a macrophage-like cell line). However, compared to the parent, the fkpA mutant survived less well in both types of cells during the first 6 h after infection. The number of viable intracellular S. typhimurium Copenhagen bacteria remained constant 6 h after infection of Caco-2 cells, but the viability of S. typhimurium KY32H1 decreased significantly by 4 h postinfection. The fkpA mutant also exhibited a reduced ability to survive intracellularly in J774.A1 cells as little as 2 h postinfection. Complementation of the fkpA mutation by a plasmid-borne wild-type fkpA gene from E. coli restored the ability of S. typhimurium KY32H1 to grow normally in J774.A1 cells. Thus, expression of the mip-like fkpA gene confers on S. typhimurium Copenhagen properties analogous to those mediated by the Mip proteins in other intracellular pathogens, suggesting that this mechanism may play a role in the virulence and/or intracellular growth of numerous bacteria.
authors.affiliation: Department of Microbiology and Immunology, School of Medicine, University of North Dakota, Grand Forks 58202-9037, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Horne
authors.forename: T J
authors.initials: TJ
authors.lastname: Kottom
authors.forename: L K
authors.initials: LK
authors.lastname: Nolan
authors.forename: K D
authors.initials: KD
authors.lastname: Young
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology; D018938:Caco-2 Cells; D004848:Epithelium / Q000382:microbiology; D005798:Genes, Bacterial; D006801:Humans; D020104:Immunophilins; D007424:Intracellular Fluid / Q000382:microbiology*; D008264:Macrophages / Q000382:microbiology; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology; D051379:Mice; D016296:Mutagenesis; D019696:Peptidylprolyl Isomerase; D010641:Phenotype; D012486:Salmonella typhimurium / Q000235:genetics* / Q000254:growth & development* / Q000472:pathogenicity; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D020104:Immunophilins; C512922:Mip protein, Legionella pneumophila; D019696:Peptidylprolyl Isomerase