title: BteA Secreted from the Bordetella bronchiseptica Type III Secetion System Induces Necrosis through an Actin Cytoskeleton Signaling Pathway and Inhibits Phagocytosis by Macrophages.
abstract: BteA is one of the effectors secreted from the Bordetella bronchiseptica type III secretion system. It has been reported that BteA induces necrosis in mammalian cells; however, the roles of BteA during the infection process are largely unknown. In order to investigate the BteA functions, morphological changes of the cells infected with the wild-type B. bronchiseptica were examined by time-lapse microscopy. L2 cells, a rat lung epithelial cell line, spread at 1.6 hours after B. bronchiseptica infection. Membrane ruffles were observed at peripheral parts of infected cells during the cell spreading. BteA-dependent cytotoxicity and cell detachment were inhibited by addition of cytochalasin D, an actin polymerization inhibitor. Domain analyses of BteA suggested that two separate amino acid regions, 200-312 and 400-658, were required for the necrosis induction. In order to examine the intra/intermolecular interactions of BteA, the amino- and the carboxyl-terminal moieties were purified as recombinant proteins from Escherichia coli. The amino-terminal moiety of BteA appeared to interact with the carboxyl-terminal moiety in the pull-down assay in vitro. When we measured the amounts of bacteria phagocytosed by J774A.1, a macrophage-like cell line, the phagocytosed amounts of B. bronchiseptica strains that deliver BteA into the host cell cytoplasm were significantly lower than those of strains that lost the ability to translocate BteA into the host cell cytoplasm. These results suggest that B. bronchiseptica induce necrosis by exploiting the actin polymerization signaling pathway and inhibit macrophage phagocytosis.
authors.affiliation: Laboratory of Bacterial Infection, Graduate School of Infection Control Sciences, Kitasato University, Tokyo 108-8641, Japan.
authors.forename: Asaomi
authors.initials: A
authors.lastname: Kuwae
authors.affiliation: Laboratory of Viral Infection II, Graduate School of Infection Control Sciences, Kitasato University, Tokyo 108-8641, Japan.
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Momose
authors.affiliation: Laboratory of Bacterial Infection, Graduate School of Infection Control Sciences, Kitasato University, Tokyo 108-8641, Japan.
authors.forename: Kanna
authors.initials: K
authors.lastname: Nagamatsu
authors.affiliation: Laboratory of Bacterial Infection, Graduate School of Infection Control Sciences, Kitasato University, Tokyo 108-8641, Japan.
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Suyama
authors.affiliation: Laboratory of Bacterial Infection, Graduate School of Infection Control Sciences, Kitasato University, Tokyo 108-8641, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Abe
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D058947:Bacterial Secretion Systems / Q000187:drug effects; D016950:Bordetella bronchiseptica / Q000502:physiology*; D019556:COS Cells; D048430:Cell Shape / Q000187:drug effects; D002522:Chlorocebus aethiops; D003571:Cytochalasin B / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D005839:Gentamicins / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism* / Q000382:microbiology*; D051379:Mice; D050505:Mutant Proteins / Q000378:metabolism; D009336:Necrosis; D010586:Phagocytes / Q000378:metabolism / Q000382:microbiology; D010587:Phagocytosis / Q000187:drug effects; D055503:Protein Multimerization / Q000187:drug effects; D017434:Protein Structure, Tertiary; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D059008:Time-Lapse Imaging
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D005839:Gentamicins; D050505:Mutant Proteins; D003571:Cytochalasin B; D007770:L-Lactate Dehydrogenase
references.citation: J Bacteriol. 2004 Jun;186(12):3806-13
references.pmid: 15175294
references.citation: RNA Biol. 2015;12(2):175-85
references.pmid: 25674816
references.citation: Infect Immun. 1984 Jan;43(1):263-9
references.pmid: 6317569
references.citation: Clin Microbiol Rev. 1991 Jul;4(3):243-55
references.pmid: 1889042
references.citation: Cell. 1992 Aug 7;70(3):401-10
references.pmid: 1643658
references.citation: Science. 1998 Jan 23;279(5350):509-14
references.pmid: 9438836
references.citation: Clin Infect Dis. 1998 Feb;26(2):389-92
references.pmid: 9502460
references.citation: Mol Microbiol. 1998 Jun;28(5):945-59
references.pmid: 9663681
references.citation: Clin Microbiol Rev. 2005 Apr;18(2):326-82
references.pmid: 15831828
references.citation: Mol Microbiol. 2005 Oct;58(1):267-79
references.pmid: 16164564
references.citation: J Biol Chem. 2006 Mar 10;281(10):6589-600
references.pmid: 16407269
references.citation: Infect Immun. 2000 Mar;68(3):1289-96
references.pmid: 10678940
references.citation: J Biol Chem. 2001 Aug 24;276(34):32230-9
references.pmid: 11413141
references.citation: EMBO J. 2003 Jun 16;22(12):2959-69
references.pmid: 12805211
references.citation: Microbiology. 2003 Sep;149(Pt 9):2513-27
references.pmid: 12949176
references.citation: Cell Microbiol. 2003 Dec;5(12):973-83
references.pmid: 14641181
references.citation: J Immunol. 2008 Dec 1;181(11):7630-8
references.pmid: 19017951
references.citation: Cell Microbiol. 2009 Dec;11(12):1735-49
references.pmid: 19650828
references.citation: J Exp Med. 2009 Dec 21;206(13):3073-88
references.pmid: 20008527
references.citation: PLoS One. 2012;7(6):e38925
references.pmid: 22701731
references.citation: Nat Methods. 2012 Jul;9(7):671-5
references.pmid: 22930834
references.citation: Nat Struct Mol Biol. 2014 Jan;21(1):82-7
references.pmid: 24317488
references.citation: J Gen Microbiol. 1970 Oct;63(2):211-20
references.pmid: 4324651