Atypical roles for Campylobacter jejuni amino acid ATP binding cassette transporter components PaqP and PaqQ in bacterial stress tolerance and pathogen-host cell dynamics.
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.
Lin
Ann E
AE
The University of British Columbia, Department of Microbiology and Immunology, Life Sciences Centre, 2350 Health Sciences Mall, Vancouver, British Columbia, V6T 1Z3, Canada.
Krastel
Kirsten
K
Hobb
Rhonda I
RI
Thompson
Stuart A
SA
Cvitkovitch
Dennis G
DG
Gaynor
Erin C
EC
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*
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins
Microbiology (Reading). 2004 Mar;150(Pt 3):561-569
14993305
FEMS Microbiol Rev. 2005 Nov;29(5):1021-40
16023758
Am J Physiol Gastrointest Liver Physiol. 2005 Sep;289(3):G388-96
15878985
J Vet Med B Infect Dis Vet Public Health. 2006 Oct;53(8):351-7
17010037
Infect Immun. 2007 Sep;75(9):4541-51
17576754
Free Radic Biol Med. 1993 Jul;15(1):57-67
8103030
Science. 2000 Oct 13;290(5490):354-7
11030657
Infect Immun. 2004 Jan;72(1):260-8
14688104
EMBO J. 2003 Feb 3;22(3):427-37
12554644
Curr Microbiol. 2008 Apr;56(4):293-7
18180992
J Infect Dis. 1993 Jul;168(1):210-5
8515112
Infect Immun. 2002 Jun;70(6):2877-85
12010975
Nature. 2000 Feb 10;403(6770):665-8
10688204
Infect Immun. 2006 May;74(5):2697-705
16622206
Clin Infect Dis. 2001 Apr 15;32(8):1201-6
11283810
Mol Microbiol. 2001 Feb;39(4):960-72
11251816
Appl Environ Microbiol. 2000 Jul;66(7):3110-2
10877815
Infect Immun. 2003 May;71(5):2626-33
12704137
J Appl Microbiol. 2006 Apr;100(4):623-32
16553716
Cell Microbiol. 2008 Jan;10(1):53-66
18052944
Mol Microbiol. 2008 Apr;68(2):474-91
18284594
Cell Microbiol. 2000 Aug;2(4):265-73
11207583
Pharmacol Res. 2003 Apr;47(4):355-62
12644394
J Bacteriol. 2002 Mar;184(5):1225-33
11844750
J Biol Chem. 2006 Oct 13;281(41):31119-30
16912041
J Bacteriol. 1997 Mar;179(6):2092-5
9068663
Microbiology (Reading). 2007 Sep;153(Pt 9):2952-2963
17768238
Mol Microbiol. 2005 Jul;57(1):143-55
15948956
Microb Pathog. 2007 Nov-Dec;43(5-6):198-207
17600669
Int J Food Microbiol. 2002 Apr 5;74(3):177-88
11981968
J Biol Chem. 2000 Dec 15;275(50):39435-43
10993883
Cell Microbiol. 2003 Mar;5(3):133-42
12614457
Toxicol Appl Pharmacol. 2007 Oct 15;224(2):174-81
17651772
Infect Immun. 2000 Aug;68(8):4384-90
10899834
Mol Microbiol. 2005 Aug;57(3):735-50
16045618
Infect Immun. 1998 Mar;66(3):1106-12
9488402
Immunobiology. 2001 Dec;204(5):543-50
11846217
Cell Signal. 2001 Feb;13(2):85-94
11257452
J Bacteriol. 1993 Oct;175(20):6546-52
8407831
Infect Immun. 1998 Mar;66(3):938-43
9488379
Infect Immun. 2006 Dec;74(12):6581-9
17015453
Mol Microbiol. 2009 Jan;71(1):253-72
19017270
Microbiology (Reading). 2005 Mar;151(Pt 3):905-915
15758235
Microbiology (Reading). 2005 Aug;151(Pt 8):2765-2772
16079352
Int J Food Microbiol. 2006 Dec 1;112(3):200-7
16782221
J Bacteriol. 1993 Sep;175(18):5899-906
8376337
Microbiology (Reading). 2007 Jan;153(Pt 1):254-62
17185554
J Nutr. 2003 Oct;133(10):3065-71
14519785
J Bacteriol. 2007 Nov;189(22):8402-3
17873037
J Biol Chem. 1997 Jun 20;272(25):15920-7
9188492
Appl Environ Microbiol. 1999 Jun;65(6):2540-6
10347040
J Bacteriol. 1999 Aug;181(16):4798-804
10438747
Infect Immun. 2006 Sep;74(9):5261-71
16926420
Mol Cell Biochem. 1981 Sep 25;39:357-68
6118826
Am J Physiol Cell Physiol. 2007 Apr;292(4):C1339-52
17135301
Infect Immun. 2002 Nov;70(11):6242-50
12379703
J Biol Chem. 2004 Sep 3;279(36):37597-612
15215244
Science. 1995 Nov 24;270(5240):1326-31
7481820
Vet Microbiol. 2007 Sep 20;124(1-2):115-24
17540517
Infect Immun. 2000 Nov;68(11):6337-45
11035743
Mol Microbiol. 2004 Apr;52(2):471-84
15066034
Anat Rec (Hoboken). 2007 Feb;290(2):199-205
17441212
Mol Microbiol. 1995 Jul;17(1):155-67
7476202
Mol Microbiol. 2006 Jun;60(5):1262-75
16689801
Cell Microbiol. 2006 Dec;8(12):1877-87
16817909
Adv Exp Med Biol. 1999;473:215-24
10659361
J Bacteriol. 2007 Nov;189(22):8099-108
17827292
Infect Immun. 1998 Aug;66(8):3666-72
9673247
Mol Microbiol. 1996 Nov;22(4):655-66
8951813
J Mol Biol. 2007 Sep 7;372(1):160-71
17631313
J Bacteriol. 1999 Dec;181(24):7588-96
10601218
J Biol Chem. 1974 Dec 25;249(24):7747-55
4279250
PLoS Pathog. 2008 Jan;4(1):e14
18225954
Am J Physiol Gastrointest Liver Physiol. 2007 Dec;293(6):G1262-71
17916648
Nat Rev Microbiol. 2007 Sep;5(9):665-79
17703225
Cell Microbiol. 2005 May;7(5):655-65
15839895
Semin Respir Infect. 1986 Jun;1(2):99-106
2825313
Infect Immun. 1985 May;48(2):446-51
3988342
Cell Host Microbe. 2007 May 17;1(3):167-74
18005696
Curr Opin Struct Biol. 2002 Dec;12(6):754-60
12504680
Appl Environ Microbiol. 2005 Jul;71(7):4004-13
16000815
Microb Pathog. 2007 Aug-Sep;43(2-3):120-5
17512161
Curr Top Microbiol Immunol. 2005;289:131-50
15791954
Pediatr Res. 2005 Dec;58(6):1192-7
16306192
Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9609-10
12122215
Infect Immun. 1998 May;66(5):1934-40
9573072
Infect Immun. 2006 Aug;74(8):4694-707
16861657
Infect Immun. 2005 Aug;73(8):5194-7
16041038
Appl Environ Microbiol. 2005 Jun;71(6):3205-12
15933023
Trends Microbiol. 2007 Oct;15(10):448-55
17920277
Mol Microbiol. 2005 Apr;56(1):8-27
15773975
Cell Host Microbe. 2008 Nov 13;4(5):425-33
18996343
Clin Immunol. 2002 Aug;104(2):151-60
12165276
Mol Microbiol. 2007 Apr;64(2):396-406
17381553
Cell Microbiol. 2007 Jan;9(1):131-41
16889624