Citrobacter rodentium lifA/efa1 is essential for colonic colonization and crypt cell hyperplasia in vivo.
Previously, we have identified a large gene (lifA, for lymphocyte inhibitory factor A) in enteropathogenic Escherichia coli (EPEC) encoding a protein termed lymphostatin that suppresses cytokine expression in vitro. This protein also functions as an adhesion factor for enterohemorrhagic E. coli (EHEC) and Shiga toxin-producing E. coli and is alternatively known as efa1 (EHEC factor for adherence 1). The lifA/efa1 gene is also present in Citrobacter rodentium, an enteric pathogen that causes a disease termed transmissible murine colonic hyperplasia (TMCH), which induces colitis and massive crypt cell proliferation, in mice. To determine if lifA/efa1 is required for C. rodentium-induced colonic pathology in vivo, three in-frame mutations were generated, disrupting the glycosyltransferase (GlM12) and protease (PrMC31) motifs and a domain in between that does not encode any known activity (EID3). In contrast to infection with wild-type C. rodentium, that with any of the lifA/efa1 mutant strains did not induce weight loss or TMCH. Enteric infection with motif mutants GlM12 and PrM31 resulted in significantly reduced colonization counts during the entire 20-day course of infection. In contrast, EID3 was indistinguishable from the wild type during the initial colonic colonization, but cleared rapidly after day 8 of the infection. The colonic epithelium of all infected mice displayed increased epithelial regeneration. However, significantly increased regeneration was observed by day 20 only in mice infected with the wild-type in comparison to those infected with lifA/efa1 mutant EID3. In summary, lifA/efa1 is a critical gene outside the locus for enterocyte effacement that regulates bacterial colonization, crypt cell proliferation, and epithelial cell regeneration.
Klapproth
Jan-Michael A
JM
Division of Digestive Diseases, Suite 201, Whitehead Biomedical Research Building, Emory University, 615 Michael St., Atlanta, GA 30345-2173, USA. jklappr@emory.edu
Sasaki
Maiko
M
Sherman
Melanie
M
Babbin
Brian
B
Donnenberg
Michael S
MS
Fernandes
Paula J
PJ
Scaletsky
Isabel C A
IC
Kalman
Daniel
D
Nusrat
Asma
A
Williams
Ifor R
IR
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D044082:Citrobacter rodentium / Q000472:pathogenicity*; D003092:Colitis / Q000382:microbiology; D003106:Colon / Q000166:cytology / Q000382:microbiology* / Q000473:pathology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D006965:Hyperplasia / Q000382:microbiology; D007223:Infant; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D009154:Mutation; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D001427:Bacterial Toxins; C404907:Efa1 protein, E coli; D029968:Escherichia coli Proteins; C406587:lymphostatin protein, E coli
Infect Immun. 1999 Nov;67(11):6019-25
10531262
Infect Immun. 1999 Jun;67(6):3031-9
10338516
Mol Microbiol. 2000 Jan;35(2):275-88
10652089
Infect Immun. 2000 Apr;68(4):2148-55
10722613
Int Immunol. 2000 May;12(5):691-700
10784615
Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5
10829079
Infect Immun. 2000 Nov;68(11):6378-83
11035748
J Clin Microbiol. 2000 Dec;38(12):4343-50
11101562
Infect Immun. 2001 Mar;69(3):1298-305
11179291
EMBO J. 2001 Mar 15;20(6):1245-58
11250891
Infect Immun. 2001 Apr;69(4):2107-15
11254564
Cancer Res. 2001 Apr 1;61(7):2899-906
11306465
Infect Immun. 2001 Oct;69(10):6217-24
11553563
Infect Immun. 2001 Oct;69(10):6323-35
11553577
Infect Immun. 2001 Nov;69(11):6660-9
11598035
Cell. 2002 May 31;109(5):575-88
12062101
Infect Immun. 2002 Sep;70(9):5158-66
12183566
Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):904-9
12538863
Mol Microbiol. 2003 Apr;48(1):95-115
12657048
Nucleic Acids Res. 2003 Apr 15;31(8):2134-47
12682364
Mol Microbiol. 2003 May;48(3):795-809
12694622
Microb Pathog. 2003 May;34(5):205-15
12732469
J Immunol. 2004 Jan 15;172(2):1146-56
14707090
Mol Microbiol. 2004 Mar;51(5):1233-49
14982621
Lancet. 1978 May 27;1(8074):1119-22
77415
Infect Immun. 1983 Sep;41(3):1340-51
6350186
J Clin Microbiol. 1985 Nov;22(5):838-40
3902881
J Bacteriol. 1991 Jan;173(1):86-93
1846140
Infect Immun. 1993 Jun;61(6):2486-92
8500884
J Clin Invest. 1993 Sep;92(3):1418-24
8376595
Nucleic Acids Res. 1994 Jun 25;22(12):2392-8
8036169
Infect Immun. 1995 Jun;63(6):2248-54
7768605
Nature. 1995 Jun 8;375(6531):500-3
7777059
J Biol Chem. 1996 Apr 26;271(17):10149-53
8626575
Mol Microbiol. 1996 Apr;20(2):313-23
8733230
Mol Microbiol. 1996 Apr;20(2):325-37
8733231
J Clin Microbiol. 1996 Oct;34(10):2364-7
8880480
Am J Pathol. 1996 Nov;149(5):1553-64
8909245
Mol Microbiol. 1997 Jan;23(2):399-407
9044273
Infect Immun. 1998 Jan;66(1):52-8
9423838
J Biol Chem. 1998 Jul 31;273(31):19566-72
9677381
J Clin Invest. 1998 Oct 15;102(8):1473-80
9788959
Science. 1999 Jul 23;285(5427):588-91
10417389
Infect Immun. 1999 Sep;67(9):4834-42
10456938
Science. 1999 Oct 15;286(5439):525-8
10521347
Am J Physiol. 1999 Apr;276(4 Pt 1):G941-50
10198338