title: Recombination-based in vivo expression technology identifies Helicobacter pylori genes important for host colonization.
abstract: Here we undertook to identify colonization and gastric disease-promoting factors of the human gastric pathogen Helicobacter pylori as genes that were induced in response to the stomach environment. Using recombination-based in vivo expression technology (RIVET), we identified six promoters induced in the host compared to laboratory conditions. Three of these promoters, designated Pivi10, Pivi66, and Pivi77, regulate genes that H. pylori may use to interact with other microbes or the host. Pivi10 likely regulates the mobA, mobB, and mobD genes, which have potential roles in horizontal gene transfer through plasmid mobilization. Pivi66 occurs in the cytotoxin-associated gene pathogenicity island, a genomic region known to be associated with more severe disease outcomes, and likely regulates cagZ, virB11, and virD4. Pivi77 likely regulates HP0289, an uncharacterized paralogue of the vacA cytotoxin gene. We assessed the roles of a subset of these genes in colonization by creating deletion mutants and analyzing them in single-strain and coinfection experiments. We found that a mobABD mutant was defective for murine host colonization and that a cagZ mutant outcompeted the wild-type strain in a coinfection analysis. Our work supports the conclusion that RIVET is a valuable tool for identifying H. pylori factors with roles in host colonization.
authors.affiliation: Department of Environmental Toxicology, University of California, Santa Cruz, 1156 High St., Santa Cruz, California 95064, USA. acastillo@mail.ewu.edu
authors.forename: Andrea R
authors.initials: AR
authors.lastname: Castillo
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Woodruff
authors.forename: Lynn E
authors.initials: LE
authors.lastname: Connolly
authors.forename: William E
authors.initials: WE
authors.lastname: Sause
authors.forename: Karen M
authors.initials: KM
authors.lastname: Ottemann
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D005798:Genes, Bacterial; D016481:Helicobacter Infections / Q000235:genetics; D016480:Helicobacter pylori / Q000235:genetics* / Q000472:pathogenicity*; D051379:Mice; D011401:Promoter Regions, Genetic
references.citation: Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5791-5
references.pmid: 8516329
references.citation: J Bacteriol. 2003 Jul;185(13):3764-72
references.pmid: 12813069
references.citation: EMBO J. 1999 Oct 15;18(20):5517-27
references.pmid: 10523296
references.citation: N Engl J Med. 2001 Sep 13;345(11):784-9
references.pmid: 11556297
references.citation: J Bacteriol. 2006 Jul;188(13):4787-800
references.pmid: 16788188
references.citation: Proteins. 1994 Dec;20(4):347-55
references.pmid: 7731953
references.citation: Mol Microbiol. 2002 Jul;45(1):1-8
references.pmid: 12100543
references.citation: J Exp Med. 2003 Apr 7;197(7):813-22
references.pmid: 12668646
references.citation: Mol Microbiol. 2001 Jul;41(2):379-91
references.pmid: 11489125
references.citation: Cancer Sci. 2005 Dec;96(12):835-43
references.pmid: 16367902
references.citation: Aliment Pharmacol Ther. 1995;9 Suppl 2:45-51
references.pmid: 8547528
references.citation: Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3516-20
references.pmid: 7724592
references.citation: Infect Immun. 2001 Nov;69(11):6769-75
references.pmid: 11598049
references.citation: Infect Immun. 2005 Feb;73(2):972-80
references.pmid: 15664940
references.citation: Mol Microbiol. 2001 Dec;42(5):1337-48
references.pmid: 11886563
references.citation: Mol Microbiol. 2005 Feb;55(3):788-98
references.pmid: 15661004
references.citation: Gastroenterology. 1992 Nov;103(5):1580-6
references.pmid: 1426878
references.citation: FEMS Microbiol Lett. 2003 Jul 15;224(1):1-15
references.pmid: 12855161
references.citation: Science. 1993 Jan 29;259(5095):686-8
references.pmid: 8430319
references.citation: Infect Immun. 2005 Feb;73(2):803-11
references.pmid: 15664919
references.citation: Infect Immun. 2001 Feb;69(2):730-6
references.pmid: 11159961
references.citation: EMBO J. 1991 Nov;10(11):3541-8
references.pmid: 1655422
references.citation: Infect Immun. 2007 Feb;75(2):1005-16
references.pmid: 17101654
references.citation: Trends Microbiol. 2000 Mar;8(3):128-33
references.pmid: 10707066
references.citation: Infect Immun. 2002 Apr;70(4):1984-90
references.pmid: 11895962
references.citation: J Bacteriol. 2001 Feb;183(4):1259-68
references.pmid: 11157938
references.citation: Mol Microbiol. 2002 Mar;43(5):1359-65
references.pmid: 11918819
references.citation: Mol Microbiol. 2003 Jul;49(1):219-34
references.pmid: 12823823
references.citation: J Clin Microbiol. 2007 Feb;45(2):402-8
references.pmid: 17135441
references.citation: J Exp Med. 2000 Feb 21;191(4):587-92
references.pmid: 10684850
references.citation: J Infect Dis. 1988 Apr;157(4):850-1
references.pmid: 3346576
references.citation: Cell. 1999 Dec 10;99(6):625-34
references.pmid: 10612398
references.citation: Nat Genet. 2001 Dec;29(4):375-6
references.pmid: 11726922
references.citation: Infect Immun. 2000 Apr;68(4):2016-23
references.pmid: 10722597
references.citation: FEMS Microbiol Lett. 2007 Sep;274(2):163-72
references.pmid: 17610513
references.citation: J Exp Med. 2000 May 1;191(9):1467-76
references.pmid: 10790422
references.citation: Proc Natl Acad Sci U S A. 1996 Feb 6;93(3):1259-64
references.pmid: 8577751
references.citation: J Infect Dis. 2004 Sep 1;190(5):946-56
references.pmid: 15295700
references.citation: J Bacteriol. 2004 Dec;186(23):7926-35
references.pmid: 15547264
references.citation: Proc Natl Acad Sci U S A. 1995 May 9;92(10):4532-6
references.pmid: 7753838
references.citation: Nat Rev Mol Cell Biol. 2001 Jun;2(6):457-66
references.pmid: 11389469
references.citation: C R Biol. 2004 Jun;327(6):551-5
references.pmid: 15330254
references.citation: Mol Microbiol. 2004 Dec;54(5):1199-211
references.pmid: 15554962
references.citation: Nat Struct Biol. 2001 Dec;8(12):1031-6
references.pmid: 11685226
references.citation: Eur J Biochem. 1991 Nov 1;201(3):653-9
references.pmid: 1718749
references.citation: J Bacteriol. 2004 Sep;186(17):5721-9
references.pmid: 15317777
references.citation: FEMS Microbiol Rev. 2004 Feb;28(1):79-100
references.pmid: 14975531
references.citation: Gastroenterology. 1997 Apr;112(4):1386-97
references.pmid: 9098027
references.citation: Science. 1998 Jan 16;279(5349):373-7
references.pmid: 9430586
references.citation: Nature. 1997 Aug 7;388(6642):539-47
references.pmid: 9252185
references.citation: Infect Immun. 2003 Mar;71(3):1379-88
references.pmid: 12595455
references.citation: Cell Microbiol. 2000 Apr;2(2):155-64
references.pmid: 11207572
references.citation: Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2634-8
references.pmid: 8146167
references.citation: EMBO J. 2003 May 1;22(9):1969-80
references.pmid: 12727865
references.citation: Mol Microbiol. 1995 Nov;18(4):671-83
references.pmid: 8817490
references.citation: Microbiology (Reading). 2002 May;148(Pt 5):1447-56
references.pmid: 11988519
references.citation: J Mol Biol. 2004 Jul 16;340(4):881-9
references.pmid: 15223328
references.citation: Mol Microbiol. 1998 Mar;27(5):967-76
references.pmid: 9535086
references.citation: J Biol Chem. 1999 Dec 31;274(53):38071-82
references.pmid: 10608876
references.citation: Infect Immun. 1996 Jul;64(7):2445-8
references.pmid: 8698465
references.citation: Mol Microbiol. 2008 Jan;67(1):155-70
references.pmid: 18078442
references.citation: Mol Cell. 2003 Sep;12(3):689-98
references.pmid: 14527414
references.citation: J Mol Biol. 1998 May 29;279(1):175-88
references.pmid: 9636708
references.citation: Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14648-53
references.pmid: 8962108
references.citation: Microbiology (Reading). 1999 Aug;145 ( Pt 8):2043-2050
references.pmid: 10463170
references.citation: Microbiol Mol Biol Rev. 1999 Sep;63(3):642-74
references.pmid: 10477311
references.citation: J Bacteriol. 1975 Dec;124(3):1502-7
references.pmid: 1104589
references.citation: FEMS Immunol Med Microbiol. 2002 Feb 18;32(3):219-26
references.pmid: 11934567
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 27;103(26):9999-10004
references.pmid: 16788065
references.citation: Appl Environ Microbiol. 2008 Apr;74(7):2095-102
references.pmid: 18245237
references.citation: Nature. 1999 Jan 14;397(6715):176-80
references.pmid: 9923682
references.citation: Science. 2000 Feb 25;287(5457):1497-500
references.pmid: 10688800
references.citation: J Bacteriol. 1996 Nov;178(21):6151-7
references.pmid: 8892813
references.citation: Proc Natl Acad Sci U S A. 2007 Apr 24;104(17):7235-40
references.pmid: 17438279
references.citation: Res Microbiol. 2005 Jan-Feb;156(1):1-6
references.pmid: 15636742
references.citation: J Biol Chem. 1997 Oct 3;272(40):25339-44
references.pmid: 9312153
references.citation: EMBO J. 1995 Jul 3;14(13):3206-14
references.pmid: 7621833
references.citation: Microbiology (Reading). 2005 Nov;151(Pt 11):3493-3503
references.pmid: 16272373
references.citation: Cell. 1984 Sep;38(2):463-9
references.pmid: 6088082
references.citation: Infect Immun. 2005 Aug;73(8):4895-904
references.pmid: 16041003
references.citation: Gastroenterology. 2002 Nov;123(5):1637-48
references.pmid: 12404238
references.citation: Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10861-5
references.pmid: 8248182
references.citation: N Engl J Med. 1991 Oct 17;325(16):1127-31
references.pmid: 1891020
references.citation: Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2001-6
references.pmid: 10051584
references.citation: Infect Immun. 2004 Apr;72(4):2358-68
references.pmid: 15039361
references.citation: J Bacteriol. 2000 Jul;182(14):3885-95
references.pmid: 10869063
references.citation: J Clin Invest. 2004 Feb;113(3):321-33
references.pmid: 14755326
references.citation: Curr Opin Microbiol. 2006 Apr;9(2):207-17
references.pmid: 16529981
references.citation: J Bacteriol. 2000 Jul;182(14):3948-54
references.pmid: 10869072
references.citation: Proc Natl Acad Sci U S A. 2000 Jun 20;97(13):7533-8
references.pmid: 10852959
references.citation: J Bacteriol. 2002 May;184(10):2755-66
references.pmid: 11976306
references.citation: Science. 2002 Jul 26;297(5581):573-8
references.pmid: 12142529