title: Citrobacter rodentium lifA/efa1 is essential for colonic colonization and crypt cell hyperplasia in vivo.
abstract: 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.
authors.affiliation: Division of Digestive Diseases, Suite 201, Whitehead Biomedical Research Building, Emory University, 615 Michael St., Atlanta, GA 30345-2173, USA. jklappr@emory.edu
authors.forename: Jan-Michael A
authors.initials: JM
authors.lastname: Klapproth
authors.forename: Maiko
authors.initials: M
authors.lastname: Sasaki
authors.forename: Melanie
authors.initials: M
authors.lastname: Sherman
authors.forename: Brian
authors.initials: B
authors.lastname: Babbin
authors.forename: Michael S
authors.initials: MS
authors.lastname: Donnenberg
authors.forename: Paula J
authors.initials: PJ
authors.lastname: Fernandes
authors.forename: Isabel C A
authors.initials: IC
authors.lastname: Scaletsky
authors.forename: Daniel
authors.initials: D
authors.lastname: Kalman
authors.forename: Asma
authors.initials: A
authors.lastname: Nusrat
authors.forename: Ifor R
authors.initials: IR
authors.lastname: Williams
mesh_terms: 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
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; C404907:Efa1 protein, E coli; D029968:Escherichia coli Proteins; C406587:lymphostatin protein, E coli
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