title: Functional impact of endotoxin receptor CD14 polymorphisms on transcriptional activity.
abstract: The polymorphism rs2569190 within the CD14 endotoxin (lipopolysaccharide, LPS) receptor gene is associated with various disease conditions that are assumed to rely on endotoxin sensitivity. In vitro experiments suggest that the T allele sensitizes the host for exogenous or endogenous LPS via an enhanced CD14 expression. To prove the impact of this single nucleotide polymorphism in its natural genomic context in vivo, two parameters of gene transcription were analyzed in peripheral blood mononuclear cells (PBMC) from single healthy individuals: (a) recruitment of RNA polymerase II by haplotype-specific chromatin immunoprecipitation and (b) the relative amount of transcripts by allele-specific transcript quantification (ASTQ). RNA polymerase II was found to be twice as much bound to the most prevalent haplotype, C-T-C-G, the only one carrying a T at the position rs2569190 of interest. ASTQ employing two independent read-out assays revealed, however, similar transcript numbers originating from C-T-C-G and non-C-T-C-G haplotypes. Total CD14 mRNA levels from freshly isolated PBMC, moreover, were neither related to donors' geno- nor haplogenotypes. Our data argue for a functional impact of the rs2569190 polymorphism in terms of a stronger transcription initiation on T allele gene variants even if preferential allele-specific binding does not result in an increase in transcript numbers. Endotoxin sensitivity associated with this genetic variation appears not to rely solely on a cis-acting regulatory impact of rs2569190 on CD14 gene transcription in PBMC.
authors.affiliation: Department of Gastroenterology and Endocrinology, University Medical Center Göttingen, 37075 Göttingen, Germany.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Mertens
authors.forename: Rusudan
authors.initials: R
authors.lastname: Bregadze
authors.forename: Ashham
authors.initials: A
authors.lastname: Mansur
authors.forename: Eva
authors.initials: E
authors.lastname: Askar
authors.forename: Heike
authors.initials: H
authors.lastname: Bickeböller
authors.forename: Giuliano
authors.initials: G
authors.lastname: Ramadori
authors.forename: Sabine
authors.initials: S
authors.lastname: Mihm
mesh_terms: D000328:Adult; D000483:Alleles; D005260:Female; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D012333:RNA, Messenger / Q000235:genetics; D011971:Receptors, Immunologic / Q000235:genetics*; D015533:Transcriptional Activation; D055815:Young Adult
chemical_list: D018950:Lipopolysaccharide Receptors; D012333:RNA, Messenger; D011971:Receptors, Immunologic; C048071:endotoxin receptor