title: Atypical roles for Campylobacter jejuni amino acid ATP binding cassette transporter components PaqP and PaqQ in bacterial stress tolerance and pathogen-host cell dynamics.
abstract: Campylobacter jejuni is a human pathogen causing severe diarrheal disease; however, our understanding of the survival of C. jejuni during disease and transmission remains limited. Amino acid ATP binding cassette (AA-ABC) transporters in C. jejuni have been proposed as important pathogenesis factors. We have investigated a novel AA-ABC transporter system, encoded by cj0467 to cj0469, by generating targeted deletions of cj0467 (the membrane transport component) and cj0469 (the ATPase component) in C. jejuni 81-176. The analyses described here have led us to designate these genes paqP and paqQ, respectively (pathogenesis-associated glutamine [q] ABC transporter permease [P] and ATPase [Q]). We found that loss of either component resulted in amino acid uptake defects, most notably diminished glutamine uptake. Altered resistance to a series of environmental and in vivo stresses was also observed: both mutants were hyperresistant to aerobic and organic peroxide stress, and while the DeltapaqP mutant was also hyperresistant to heat and osmotic shock, the DeltapaqQ mutant was more susceptible than the wild type to the latter two stresses. The DeltapaqP and DeltapaqQ mutants also displayed a surprising but statistically significant increase in recovery from macrophages and epithelial cells in short-term intracellular survival assays. Annexin V, 4',6-diamidino-2-phenylindole (DAPI), and Western blot analyses revealed that macrophages infected with the DeltapaqP or DeltapaqQ mutant exhibited transient but significant decreases in cell death and extracellular signal-regulated kinase-mitogen-activated protein kinase activation compared to levels in wild-type-infected cells. The DeltapaqP mutant was not defective in either short-term or longer-term mouse colonization, consistent with its increased stress survival and diminished host cell damage phenotypes. Collectively, these results demonstrate a unique correlation of an AA-ABC transporter with bacterial stress tolerances and host cell responses to pathogen infection.
authors.affiliation: The University of British Columbia, Department of Microbiology and Immunology, Life Sciences Centre, 2350 Health Sciences Mall, Vancouver, British Columbia, V6T 1Z3, Canada.
authors.forename: Ann E
authors.initials: AE
authors.lastname: Lin
authors.forename: Kirsten
authors.initials: K
authors.lastname: Krastel
authors.forename: Rhonda I
authors.initials: RI
authors.lastname: Hobb
authors.forename: Stuart A
authors.initials: SA
authors.lastname: Thompson
authors.forename: Dennis G
authors.initials: DG
authors.lastname: Cvitkovitch
authors.forename: Erin C
authors.initials: EC
authors.lastname: Gaynor
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015139:Blotting, Southern; D015153:Blotting, Western; D002169:Campylobacter Infections / Q000235:genetics / Q000378:metabolism; D016123:Campylobacter jejuni / Q000472:pathogenicity / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics; D006790:Host-Parasite Interactions / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013312:Stress, Physiological / Q000502:physiology*
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins
references.citation: Microbiology (Reading). 2004 Mar;150(Pt 3):561-569
references.pmid: 14993305
references.citation: FEMS Microbiol Rev. 2005 Nov;29(5):1021-40
references.pmid: 16023758
references.citation: Am J Physiol Gastrointest Liver Physiol. 2005 Sep;289(3):G388-96
references.pmid: 15878985
references.citation: J Vet Med B Infect Dis Vet Public Health. 2006 Oct;53(8):351-7
references.pmid: 17010037
references.citation: Infect Immun. 2007 Sep;75(9):4541-51
references.pmid: 17576754
references.citation: Free Radic Biol Med. 1993 Jul;15(1):57-67
references.pmid: 8103030
references.citation: Science. 2000 Oct 13;290(5490):354-7
references.pmid: 11030657
references.citation: Infect Immun. 2004 Jan;72(1):260-8
references.pmid: 14688104
references.citation: EMBO J. 2003 Feb 3;22(3):427-37
references.pmid: 12554644
references.citation: Curr Microbiol. 2008 Apr;56(4):293-7
references.pmid: 18180992
references.citation: J Infect Dis. 1993 Jul;168(1):210-5
references.pmid: 8515112
references.citation: Infect Immun. 2002 Jun;70(6):2877-85
references.pmid: 12010975
references.citation: Nature. 2000 Feb 10;403(6770):665-8
references.pmid: 10688204
references.citation: Infect Immun. 2006 May;74(5):2697-705
references.pmid: 16622206
references.citation: Clin Infect Dis. 2001 Apr 15;32(8):1201-6
references.pmid: 11283810
references.citation: Mol Microbiol. 2001 Feb;39(4):960-72
references.pmid: 11251816
references.citation: Appl Environ Microbiol. 2000 Jul;66(7):3110-2
references.pmid: 10877815
references.citation: Infect Immun. 2003 May;71(5):2626-33
references.pmid: 12704137
references.citation: J Appl Microbiol. 2006 Apr;100(4):623-32
references.pmid: 16553716
references.citation: Cell Microbiol. 2008 Jan;10(1):53-66
references.pmid: 18052944
references.citation: Mol Microbiol. 2008 Apr;68(2):474-91
references.pmid: 18284594
references.citation: Cell Microbiol. 2000 Aug;2(4):265-73
references.pmid: 11207583
references.citation: Pharmacol Res. 2003 Apr;47(4):355-62
references.pmid: 12644394
references.citation: J Bacteriol. 2002 Mar;184(5):1225-33
references.pmid: 11844750
references.citation: J Biol Chem. 2006 Oct 13;281(41):31119-30
references.pmid: 16912041
references.citation: J Bacteriol. 1997 Mar;179(6):2092-5
references.pmid: 9068663
references.citation: Microbiology (Reading). 2007 Sep;153(Pt 9):2952-2963
references.pmid: 17768238
references.citation: Mol Microbiol. 2005 Jul;57(1):143-55
references.pmid: 15948956
references.citation: Microb Pathog. 2007 Nov-Dec;43(5-6):198-207
references.pmid: 17600669
references.citation: Int J Food Microbiol. 2002 Apr 5;74(3):177-88
references.pmid: 11981968
references.citation: J Biol Chem. 2000 Dec 15;275(50):39435-43
references.pmid: 10993883
references.citation: Cell Microbiol. 2003 Mar;5(3):133-42
references.pmid: 12614457
references.citation: Toxicol Appl Pharmacol. 2007 Oct 15;224(2):174-81
references.pmid: 17651772
references.citation: Infect Immun. 2000 Aug;68(8):4384-90
references.pmid: 10899834
references.citation: Mol Microbiol. 2005 Aug;57(3):735-50
references.pmid: 16045618
references.citation: Infect Immun. 1998 Mar;66(3):1106-12
references.pmid: 9488402
references.citation: Immunobiology. 2001 Dec;204(5):543-50
references.pmid: 11846217
references.citation: Cell Signal. 2001 Feb;13(2):85-94
references.pmid: 11257452
references.citation: J Bacteriol. 1993 Oct;175(20):6546-52
references.pmid: 8407831
references.citation: Infect Immun. 1998 Mar;66(3):938-43
references.pmid: 9488379
references.citation: Infect Immun. 2006 Dec;74(12):6581-9
references.pmid: 17015453
references.citation: Mol Microbiol. 2009 Jan;71(1):253-72
references.pmid: 19017270
references.citation: Microbiology (Reading). 2005 Mar;151(Pt 3):905-915
references.pmid: 15758235
references.citation: Microbiology (Reading). 2005 Aug;151(Pt 8):2765-2772
references.pmid: 16079352
references.citation: Int J Food Microbiol. 2006 Dec 1;112(3):200-7
references.pmid: 16782221
references.citation: J Bacteriol. 1993 Sep;175(18):5899-906
references.pmid: 8376337
references.citation: Microbiology (Reading). 2007 Jan;153(Pt 1):254-62
references.pmid: 17185554
references.citation: J Nutr. 2003 Oct;133(10):3065-71
references.pmid: 14519785
references.citation: J Bacteriol. 2007 Nov;189(22):8402-3
references.pmid: 17873037
references.citation: J Biol Chem. 1997 Jun 20;272(25):15920-7
references.pmid: 9188492
references.citation: Appl Environ Microbiol. 1999 Jun;65(6):2540-6
references.pmid: 10347040
references.citation: J Bacteriol. 1999 Aug;181(16):4798-804
references.pmid: 10438747
references.citation: Infect Immun. 2006 Sep;74(9):5261-71
references.pmid: 16926420
references.citation: Mol Cell Biochem. 1981 Sep 25;39:357-68
references.pmid: 6118826
references.citation: Am J Physiol Cell Physiol. 2007 Apr;292(4):C1339-52
references.pmid: 17135301
references.citation: Infect Immun. 2002 Nov;70(11):6242-50
references.pmid: 12379703
references.citation: J Biol Chem. 2004 Sep 3;279(36):37597-612
references.pmid: 15215244
references.citation: Science. 1995 Nov 24;270(5240):1326-31
references.pmid: 7481820
references.citation: Vet Microbiol. 2007 Sep 20;124(1-2):115-24
references.pmid: 17540517
references.citation: Infect Immun. 2000 Nov;68(11):6337-45
references.pmid: 11035743
references.citation: Mol Microbiol. 2004 Apr;52(2):471-84
references.pmid: 15066034
references.citation: Anat Rec (Hoboken). 2007 Feb;290(2):199-205
references.pmid: 17441212
references.citation: Mol Microbiol. 1995 Jul;17(1):155-67
references.pmid: 7476202
references.citation: Mol Microbiol. 2006 Jun;60(5):1262-75
references.pmid: 16689801
references.citation: Cell Microbiol. 2006 Dec;8(12):1877-87
references.pmid: 16817909
references.citation: Adv Exp Med Biol. 1999;473:215-24
references.pmid: 10659361
references.citation: J Bacteriol. 2007 Nov;189(22):8099-108
references.pmid: 17827292
references.citation: Infect Immun. 1998 Aug;66(8):3666-72
references.pmid: 9673247
references.citation: Mol Microbiol. 1996 Nov;22(4):655-66
references.pmid: 8951813
references.citation: J Mol Biol. 2007 Sep 7;372(1):160-71
references.pmid: 17631313
references.citation: J Bacteriol. 1999 Dec;181(24):7588-96
references.pmid: 10601218
references.citation: J Biol Chem. 1974 Dec 25;249(24):7747-55
references.pmid: 4279250
references.citation: PLoS Pathog. 2008 Jan;4(1):e14
references.pmid: 18225954
references.citation: Am J Physiol Gastrointest Liver Physiol. 2007 Dec;293(6):G1262-71
references.pmid: 17916648
references.citation: Nat Rev Microbiol. 2007 Sep;5(9):665-79
references.pmid: 17703225
references.citation: Cell Microbiol. 2005 May;7(5):655-65
references.pmid: 15839895
references.citation: Semin Respir Infect. 1986 Jun;1(2):99-106
references.pmid: 2825313
references.citation: Infect Immun. 1985 May;48(2):446-51
references.pmid: 3988342
references.citation: Cell Host Microbe. 2007 May 17;1(3):167-74
references.pmid: 18005696
references.citation: Curr Opin Struct Biol. 2002 Dec;12(6):754-60
references.pmid: 12504680
references.citation: Appl Environ Microbiol. 2005 Jul;71(7):4004-13
references.pmid: 16000815
references.citation: Microb Pathog. 2007 Aug-Sep;43(2-3):120-5
references.pmid: 17512161
references.citation: Curr Top Microbiol Immunol. 2005;289:131-50
references.pmid: 15791954
references.citation: Pediatr Res. 2005 Dec;58(6):1192-7
references.pmid: 16306192
references.citation: Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9609-10
references.pmid: 12122215
references.citation: Infect Immun. 1998 May;66(5):1934-40
references.pmid: 9573072
references.citation: Infect Immun. 2006 Aug;74(8):4694-707
references.pmid: 16861657
references.citation: Infect Immun. 2005 Aug;73(8):5194-7
references.pmid: 16041038
references.citation: Appl Environ Microbiol. 2005 Jun;71(6):3205-12
references.pmid: 15933023
references.citation: Trends Microbiol. 2007 Oct;15(10):448-55
references.pmid: 17920277
references.citation: Mol Microbiol. 2005 Apr;56(1):8-27
references.pmid: 15773975
references.citation: Cell Host Microbe. 2008 Nov 13;4(5):425-33
references.pmid: 18996343
references.citation: Clin Immunol. 2002 Aug;104(2):151-60
references.pmid: 12165276
references.citation: Mol Microbiol. 2007 Apr;64(2):396-406
references.pmid: 17381553
references.citation: Cell Microbiol. 2007 Jan;9(1):131-41
references.pmid: 16889624