title: A phenotype at last: essential role for the Yersinia enterocolitica Ysa type III secretion system in a Drosophila melanogaster S2 cell model.
abstract: The highly pathogenic Yersinia enterocolitica strains have a chromosomally encoded type III secretion system (T3SS) that is expressed and functional in vitro only when the bacteria are cultured at 26 °C. Mutations that render this system nonfunctional are slightly attenuated in the mouse model of infection only following an oral inoculation and only at early time points postinfection. The discrepancy between the temperature required for the Ysa gene expression and the physiological temperature required for mammalian model systems has made defining the role of this T3SS challenging. Therefore, we explored the use of Drosophila S2 cells as a model system for studying Ysa function. We show here that Y. enterocolitica is capable of infecting S2 cells and replicating intracellularly to high levels, an unusual feature of this pathogen. Importantly, we show that the Ysa T3SS is required for robust intracellular replication. A secretion-deficient mutant lacking the secretin gene, ysaC, is defective in replication within S2 cells, marking the first demonstration of a pronounced Ysa-dependent virulence phenotype. Establishment of S2 cells as a model for Y. enterocolitica infection provides a versatile tool to elucidate the role of the Ysa T3SS in the life cycle of this gastrointestinal pathogen.
authors: Department of Genetics, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA. kawalker@med.unc.edu
authors: Kimberly A
authors: KA
authors: Walker
authors: Vivien I
authors: VI
authors: Maltez
authors: Joshua D
authors: JD
authors: Hall
authors: Nicholas P
authors: NP
authors: Vitko
authors: Virginia L
authors: VL
authors: Miller
mesh_terms: D000818:Animals; D058491:Bacterial Load; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D058947:Bacterial Secretion Systems; D002460:Cell Line; D003593:Cytoplasm / Q000382:microbiology; D004195:Disease Models, Animal; D004331:Drosophila melanogaster / Q000382:microbiology*; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008856:Microscopy, Fluorescence; D009154:Mutation; D010641:Phenotype; D011401:Promoter Regions, Genetic; D021381:Protein Transport; D013997:Time Factors; D015009:Yersinia Infections / Q000382:microbiology*; D015008:Yersinia enterocolitica / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems
references: Nat Protoc. 2008;3(4):606-11
references: 18388942
references: Gut. 1990 Sep;31(9):1011-5
references: 2210445
references: Infect Immun. 1997 Jun;65(6):2497-501
references: 9169800
references: Microbiol Mol Biol Rev. 1998 Dec;62(4):1315-52
references: 9841674
references: Nat Rev Microbiol. 2008 Jan;6(1):11-6
references: 18059288
references: Science. 2005 Aug 19;309(5738):1248-51
references: 16020693
references: Curr Opin Microbiol. 2009 Feb;12(1):117-24
references: 19157959
references: Curr Opin Microbiol. 2008 Jun;11(3):262-70
references: 18539520
references: Infect Immun. 2005 Sep;73(9):5961-77
references: 16113317
references: J Bacteriol. 2002 Oct;184(20):5563-71
references: 12270813
references: Mol Plant Microbe Interact. 2000 Nov;13(11):1243-50
references: 11059491
references: Plasmid. 2012 Jul;68(1):33-42
references: 22445322
references: Tissue Cell. 1977;9(4):751-66
references: 205012
references: Proc Natl Acad Sci U S A. 2012 Jul 31;109(31):12710-5
references: 22802624
references: Mol Microbiol. 2011 Jun;80(6):1420-38
references: 21488979
references: PLoS Pathog. 2008 Mar 21;4(3):e1000031
references: 18369477
references: Gene. 1993 Dec 22;136(1-2):271-5
references: 8294016
references: Infect Immun. 1988 May;56(5):1242-8
references: 2833444
references: Science. 2005 Aug 19;309(5738):1251-3
references: 16020694
references: Cell Microbiol. 2005 May;7(5):725-39
references: 15839901
references: Proc Natl Acad Sci U S A. 2005 Sep 20;102(38):13646-51
references: 16157870
references: PLoS One. 2010 Jun 11;5(6):e11025
references: 20552012
references: Mol Microbiol. 2000 Jun;36(6):1436-46
references: 10931293
references: PLoS Genet. 2006 Dec 15;2(12):e206
references: 17173484
references: J Bacteriol. 2002 Mar;184(5):1324-34
references: 11844761
references: J Bacteriol. 1988 Jun;170(6):2575-83
references: 2836362
references: J Cell Biol. 2002 Aug 5;158(3):401-8
references: 12163464
references: Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16669-74
references: 17925449
references: Arch Biochem Biophys. 1969 Jun;132(1):91-8
references: 4389415
references: Environ Microbiol. 2009 Jun;11(6):1473-81
references: 19220402
references: Eur J Cell Biol. 2000 Oct;79(10):659-71
references: 11089914
references: J Antimicrob Chemother. 2007 Feb;59(2):246-53
references: 17220162
references: PLoS Pathog. 2007 Oct 26;3(10):1446-58
references: 17967059
references: Cell Microbiol. 2003 Dec;5(12):875-85
references: 14641173
references: Cell Microbiol. 2009 Nov;11(11):1579-86
references: 19775254
references: Infect Immun. 1998 Mar;66(3):1237-43
references: 9488419
references: J Bacteriol. 2009 Mar;191(6):1816-26
references: 19124573
references: J Bacteriol. 2008 Nov;190(22):7315-25
references: 18805975
references: J Cell Biol. 1963 Apr;17:208-12
references: 13986422
references: Nature. 2002 Apr 11;416(6881):644-8
references: 11912489
references: Clin Immunol. 2005 Mar;114(3):216-26
references: 15721832
references: Mol Microbiol. 2006 Jan;59(2):689-706
references: 16390460
references: J Bacteriol. 2004 Jul;186(13):4056-66
references: 15205407
references: PLoS Pathog. 2006 Apr;2(4):e34
references: 16652170