title: A specific A/T polymorphism in Western tyrosine phosphorylation B-motifs regulates Helicobacter pylori CagA epithelial cell interactions.
abstract: Helicobacter pylori persistently colonizes the human stomach, with mixed roles in human health. The CagA protein, a key host-interaction factor, is translocated by a type IV secretion system into host epithelial cells, where its EPIYA tyrosine phosphorylation motifs (TPMs) are recognized by host cell kinases, leading to multiple host cell signaling cascades. The CagA TPMs have been described as type A, B, C or D, each with a specific conserved amino acid sequence surrounding EPIYA. Database searching revealed strong non-random distribution of the B-motifs (including EPIYA and EPIYT) in Western H. pylori isolates. In silico analysis of Western H. pylori CagA sequences provided evidence that the EPIYT B-TPMs are significantly less associated with gastric cancer than the EPIYA B-TPMs. By generating and using a phosphorylated CagA B-TPM-specific antibody, we demonstrated the phosphorylated state of the CagA B-TPM EPIYT during H. pylori co-culture with host cells. We also showed that within host cells, CagA interaction with phosphoinositol 3-kinase (PI3-kinase) was B-TPM tyrosine-phosphorylation-dependent, and the recombinant CagA with EPIYT B-TPM had higher affinity to PI3-kinase and enhanced induction of AKT than the isogenic CagA with EPIYA B-TPM. Structural modeling of the CagA B-TPM motif bound to PI3-kinase indicated that the threonine residue at the pY+1 position forms a side-chain hydrogen bond to N-417 of PI3-kinase, which cannot be formed by alanine. During co-culture with AGS cells, an H. pylori strain with a CagA EPIYT B-TPM had significantly attenuated induction of interleukin-8 and hummingbird phenotype, compared to the isogenic strain with B-TPM EPIYA. These results suggest that the A/T polymorphisms could regulate CagA activity through interfering with host signaling pathways related to carcinogenesis, thus influencing cancer risk.
authors: Departments of Medicine and Microbiology, New York University School of Medicine and VA Medical Center, New York, New York, United States of America.
authors: Xue-Song
authors: XS
authors: Zhang
authors: Friedrich Alexander University Erlangen, Department of Biology, Division of Microbiology, Erlangen, Germany.
authors: Nicole
authors: N
authors: Tegtmeyer
authors: Departments of Medicine and Microbiology, New York University School of Medicine and VA Medical Center, New York, New York, United States of America.
authors: Leah
authors: L
authors: Traube
authors: Departments of Medicine and Microbiology, New York University School of Medicine and VA Medical Center, New York, New York, United States of America.
authors: Shawn
authors: S
authors: Jindal
authors: Departments of Medicine and Microbiology, New York University School of Medicine and VA Medical Center, New York, New York, United States of America.
authors: Guillermo
authors: G
authors: Perez-Perez
authors: Friedrich Alexander University Erlangen, Bioinformatics, Institute for Biochemistry, Erlangen, Germany.
authors: Heinrich
authors: H
authors: Sticht
authors: Friedrich Alexander University Erlangen, Department of Biology, Division of Microbiology, Erlangen, Germany.
authors: Steffen
authors: S
authors: Backert
authors: Departments of Medicine and Microbiology, New York University School of Medicine and VA Medical Center, New York, New York, United States of America.
authors: Martin J
authors: MJ
authors: Blaser
mesh_terms: D020816:Amino Acid Motifs; D000942:Antigens, Bacterial / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D002460:Cell Line; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000382:microbiology*; D016480:Helicobacter pylori / Q000235:genetics*; D054884:Host-Pathogen Interactions / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D010766:Phosphorylation; D020641:Polymorphism, Single Nucleotide
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; C080866:cagA protein, Helicobacter pylori
references: Trends Biochem Sci. 2003 Jun;28(6):284-93
references: 12826400
references: World J Gastroenterol. 2011 Aug 28;17(32):3672-83
references: 21990948
references: J Bacteriol. 2012 Aug;194(15):3891-903
references: 22609923
references: J Mol Med (Berl). 1999 Dec;77(12):834-46
references: 10682319
references: Nat Struct Mol Biol. 2010 Jan;17(1):130-2
references: 19966800
references: Trends Biochem Sci. 2008 Feb;33(2):80-90
references: 18182299
references: Nat Rev Drug Discov. 2014 Feb;13(2):140-56
references: 24481312
references: J Biol Chem. 2011 Sep 23;286(38):33622-31
references: 21813645
references: Mol Microbiol. 1999 May;32(3):459-70
references: 10320570
references: Nature. 1997 Aug 7;388(6642):539-47
references: 9252185
references: Nat Rev Cancer. 2004 Sep;4(9):688-94
references: 15343275
references: Clin Microbiol Rev. 2010 Oct;23(4):713-39
references: 20930071
references: Proc Natl Acad Sci U S A. 2005 Jun 28;102(26):9300-5
references: 15972330
references: J Infect Dis. 2004 Mar 1;189(5):820-7
references: 14976598
references: FEBS J. 2011 Apr;278(8):1190-202
references: 21352489
references: Nature. 2007 Feb 22;445(7130):915-8
references: 17287725
references: Nat Struct Biol. 1996 Apr;3(4):364-74
references: 8599763
references: Cancer Res. 1995 May 15;55(10):2111-5
references: 7743510
references: J Biol Chem. 2010 Jul 23;285(30):23515-26
references: 20507990
references: J Clin Microbiol. 2012 Mar;50(3):876-83
references: 22189111
references: Mol Cell Proteomics. 2008 Apr;7(4):768-84
references: 17956856
references: Cell Host Microbe. 2009 Apr 23;5(4):397-403
references: 19380118
references: J Biol Chem. 2006 Oct 27;281(43):32344-52
references: 16954210
references: J Infect Dis. 2009 Mar 1;199(5):641-51
references: 19199544
references: J Infect Dis. 2009 Apr 15;199(8):1218-21
references: 19278338
references: Oncogene. 2005 Jun 2;24(24):3886-95
references: 15856031
references: Cell Commun Signal. 2011 Dec 28;9:32
references: 22204307
references: Infect Immun. 2008 Jul;76(7):3233-40
references: 18474653
references: J Clin Invest. 2012 Apr;122(4):1553-66
references: 22378042
references: J Clin Microbiol. 2009 Apr;47(4):959-68
references: 19158258
references: J Infect Dis. 2003 Aug 15;188(4):486-96
references: 12898434
references: Microbes Infect. 2011 Sep;13(10):799-807
references: 21477660
references: J Biol Chem. 1994 Feb 18;269(7):5241-8
references: 8106507
references: Cell Microbiol. 2014 Sep;16(9):1441-55
references: 24779413
references: Cell Host Microbe. 2009 Jan 22;5(1):23-34
references: 19154985
references: Gut. 1998 Sep;43(3):327-33
references: 9863476
references: PLoS One. 2011;6(8):e23499
references: 21853141
references: FEMS Microbiol Lett. 2005 Feb 1;243(1):265-70
references: 15668028
references: Microbiol Res. 2009;164(2):180-90
references: 17449233
references: J Biol Chem. 2005 Mar 18;280(11):9973-9
references: 15640153
references: Infect Genet Evol. 2012 Mar;12(2):203-13
references: 22197766
references: Infect Immun. 2010 Dec;78(12):5244-51
references: 20876293
references: Nat Rev Mol Cell Biol. 2007 Jul;8(7):530-41
references: 17585314
references: Science. 2003 Mar 7;299(5612):1582-5
references: 12624269
references: Cell Host Microbe. 2014 Mar 12;15(3):306-16
references: 24629337
references: Proc Natl Acad Sci U S A. 2014 Jan 28;111(4):1562-7
references: 24474782
references: Cell Microbiol. 2006 Jan;8(1):97-106
references: 16367869
references: J Gastroenterol. 2012 Aug;47(8):936-40
references: 22722905
references: Science. 2002 Jan 25;295(5555):683-6
references: 11743164
references: PLoS One. 2013;8(8):e71220
references: 23940723
references: J Biol Chem. 2010 May 21;285(21):16042-50
references: 20348091
references: Infect Immun. 1993 May;61(5):1799-809
references: 8478069
references: PLoS One. 2014;9(5):e96488
references: 24800748
references: Mol Cell Biol. 2006 Jan;26(1):261-76
references: 16354697
references: N Engl J Med. 1994 May 5;330(18):1267-71
references: 8145781
references: Mol Microbiol. 2002 Feb;43(4):971-80
references: 11929545
references: J Clin Invest. 2012 Apr;122(4):1192-5
references: 22378039
references: J Med Microbiol. 2008 Sep;57(Pt 9):1062-7
references: 18719174
references: Lancet. 1984 Jun 16;1(8390):1311-5
references: 6145023
references: Cell Host Microbe. 2012 Jul 19;12(1):20-33
references: 22817985
references: Nature. 2007 May 17;447(7142):330-3
references: 17507984
references: Proc Natl Acad Sci U S A. 2014 Jan 28;111(4):1455-60
references: 24474772
references: Electrophoresis. 1997 Dec;18(15):2714-23
references: 9504803
references: Clin Diagn Lab Immunol. 2002 Jan;9(1):167-75
references: 11777849
references: Proc Natl Acad Sci U S A. 1999 Dec 7;96(25):14559-64
references: 10588744
references: J Clin Pathol. 1994 Jan;47(1):61-6
references: 8132812
references: J Biol Chem. 2003 Feb 7;278(6):3664-70
references: 12446738
references: J Infect Dis. 2002 Dec 1;186(11):1621-30
references: 12447739
references: Gastroenterology. 1999 Aug;117(2):342-9
references: 10419915
references: Biomed Environ Sci. 2010 Aug;23(4):273-8
references: 20934114
references: Helicobacter. 2010 Jun;15(3):163-76
references: 20557357
references: J Exp Med. 2005 Nov 7;202(9):1235-47
references: 16275761
references: Proc Natl Acad Sci U S A. 2008 Jan 22;105(3):1003-8
references: 18192401
references: J Biol Chem. 2004 Apr 23;279(17):17205-16
references: 14963045
references: Infect Immun. 2007 Mar;75(3):1203-13
references: 17194805
references: PLoS One. 2014;9(6):e97782
references: 24893039
references: Proc Natl Acad Sci U S A. 2002 Oct 29;99(22):14428-33
references: 12391297
references: Gut Microbes. 2012 Nov-Dec;3(6):566-71
references: 22895083
references: Cell Host Microbe. 2011 Jun 16;9(6):520-31
references: 21669400
references: Gastroenterology. 2007 Apr;132(4):1309-19
references: 17408661
references: Lancet. 1996 Nov 30;348(9040):1462-3
references: 8942769
references: Trends Biochem Sci. 1995 Sep;20(9):374
references: 7482707
references: J Bacteriol. 2012 Jul;194(13):3407-16
references: 22522893
references: Oncogene. 2008 Sep 18;27(41):5497-510
references: 18794884
references: J Biol Chem. 2005 Jun 17;280(24):23130-7
references: 15831497
references: J Gastroenterol. 2009;44(5):405-11
references: 19347242