title: Transcription Factor SP2 Enhanced the Expression of Cd14 in Colitis-Susceptible C3H/HeJBir.
abstract: Genetic analysis in the IL10-deficient mouse model revealed a modifier locus of experimental inflammatory bowel disease (IBD) on chromosome 18, with the allele of the strain C3H/HeJBir (C3Bir) conferring resistance and the allele of C57BL/6J (B6) conferring susceptibility. Differential Cd14 expression was associated with this background specific susceptibility to intestinal inflammation. Polymorphisms of the Cd14 promoter were found to be likely causative for strain specific expression, and Cd14-knockout mice revealed a protective role of this gene-product in experimental IBD. In this study, luciferase reporter assays confirmed an increased activity of the C3Bir derived Cd14 promoter compared to the one of B6. Promoter truncation experiments and site-directed mutagenesis in both strains resulted in reduced Cd14 promoter activity and confirmed that a central AP1 and the proximal SP1 transcription factor binding sites mediated the basal activity of the Cd14 promoter in the mouse. Moreover, a T to C exchange at position -259 replaced putative STAT1 and CDX1 sites in the Cd14 promoter from B6 by a SP2 site in C3Bir. Ablation of the Sp2 site through truncation was associated with a decreased promoter activity. Site-directed mutagenesis also demonstrated that the inactivation of SP2 led to a substantial loss of promoter activity in C3Bir. Performing electrophoretic mobility shift and supershift assays demonstrated interaction of SP2 with its potential binding site. In addition, retroviral-mediated overexpression of the SP2 transcription factor in primary bone marrow macrophages derived from C3Bir mice caused a significant increase in Cd14 transcription. These data characterized SP2 as important factor responsible for higher Cd14 expression and reduced IBD susceptibility mediated by the C3Bir allele.
authors.affiliation: Institute for Laboratory Animal Science and Central Animal Facility, Hannover Medical School, Hannover, Germany.
authors.forename: Nils-Holger
authors.initials: NH
authors.lastname: Zschemisch
authors.affiliation: Institute for Laboratory Animal Science and Central Animal Facility, Hannover Medical School, Hannover, Germany.
authors.forename: Inga
authors.initials: I
authors.lastname: Brüsch
authors.affiliation: Institute for Laboratory Animal Science and Central Animal Facility, Hannover Medical School, Hannover, Germany.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Hambusch
authors.affiliation: Institute for Laboratory Animal Science and Central Animal Facility, Hannover Medical School, Hannover, Germany.
authors.forename: André
authors.initials: A
authors.lastname: Bleich
mesh_terms: D000483:Alleles; D000818:Animals; D002460:Cell Line; D003093:Colitis, Ulcerative / Q000235:genetics / Q000378:metabolism*; D018950:Lipopolysaccharide Receptors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051704:Sp2 Transcription Factor / Q000235:genetics / Q000378:metabolism*
chemical_list: D018950:Lipopolysaccharide Receptors; D051704:Sp2 Transcription Factor