title: A 3'UTR polymorphism marks differential KLRG1 mRNA levels through disruption of a miR-584-5p binding site and associates with pemphigus foliaceus susceptibility.
abstract: Genetic variations mapping to 3' untranslated regions (3'UTRs) may overlap with microRNA (miRNA) binding sites, therefore potentially interfering with translation inhibition or messenger RNA (mRNA) degradation. The aim of this study was to investigate whether single nucleotide polymorphisms (SNPs) located within the 3'UTRs of six candidate genes and predicted to interfere with miRNA ligation could account for disease-relevant differential mRNA levels. Focusing on pemphigus foliaceus (PF) - an autoimmune blistering skin condition with unique endemic patterns - we investigated whether nine 3'UTR SNPs from the CD1D, CTLA4, KLRD1, KLRG1, NKG7, and TNFSF13B genes differentially expressed in PF were disease-associated. The heterozygous genotype of the KLRG1 rs1805672 polymorphism was associated with increased predisposition to PF (A/G vs. A/A: P=0.038; OR=1.60), and a trend for augmented susceptibility was observed for carriers of the G allele (P=0.094; OR=1.44). In silico analyses suggested that rs1805672 G allele could disrupt binding of miR-584-5p, and indicated rs1805672 as an expression Quantitative Trait Locus (eQTL), with an effect on KLRG1 gene expression. Dual-luciferase assay showed that miR-584-5p mediated approximately 50% downregulation of the reporter gene's activity through the 3'UTR of KLRG1 harboring rs1805672 A allele (vs. miRNA-negative condition, P=0.006). This silencing relationship was lost after site-directed mutation to G allele (vs. miRNA-negative condition, P=0.391; vs. rs1805672 A allele, P=0.005). Collectively, these results suggest that a disease-associated SNP located within the 3'UTR of KLRG1 directly interferes with miR-584-5p binding, allowing for KLRG1 mRNA differential accumulation, which in turn may contribute to pathogenesis of autoimmune diseases, such as pemphigus.
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil; CAPES Foundation, Ministry of Education of Brazil, Brasília, DF 70040-020, Brazil.
authors.forename: Gabriel A
authors.initials: GA
authors.lastname: Cipolla
authors.affiliation: Deparment of Dermatology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, United States.
authors.forename: Jong Kook
authors.initials: JK
authors.lastname: Park
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil.
authors.forename: Liana A
authors.initials: LA
authors.lastname: de Oliveira
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil.
authors.forename: Sara Cristina
authors.initials: SC
authors.lastname: Lobo-Alves
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil.
authors.forename: Rodrigo C
authors.initials: RC
authors.lastname: de Almeida
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil.
authors.forename: Ticiana D J
authors.initials: TD
authors.lastname: Farias
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil.
authors.forename: Débora de S
authors.initials: Dde S
authors.lastname: Lemos
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil.
authors.forename: Danielle
authors.initials: D
authors.lastname: Malheiros
authors.affiliation: Deparment of Dermatology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, United States.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Lavker
authors.affiliation: Department of Genetics, Federal University of Paraná, Curitiba, PR 81531-980, Brazil. Electronic address: perler@ufpr.br.
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Petzl-Erler
mesh_terms: D020413:3' Untranslated Regions; D000483:Alleles; D055610:Antigens, CD1d / Q000235:genetics / Q000378:metabolism; D053264:B-Cell Activating Factor / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D060908:CTLA-4 Antigen / Q000235:genetics / Q000378:metabolism; D016022:Case-Control Studies; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D006239:Haplotypes; D006801:Humans; D037181:Lectins, C-Type / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D051920:NK Cell Lectin-Like Receptor Subfamily D / Q000235:genetics / Q000378:metabolism; D010392:Pemphigus / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D020641:Polymorphism, Single Nucleotide; D011971:Receptors, Immunologic; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; D055610:Antigens, CD1d; D053264:B-Cell Activating Factor; C526877:CD1D protein, human; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C497869:KLRD1 protein, human; C113413:KLRG1 protein, human; D037181:Lectins, C-Type; C552781:MIRN584 microRNA, human; D008565:Membrane Proteins; D035683:MicroRNAs; D051920:NK Cell Lectin-Like Receptor Subfamily D; C499363:NKG7 protein, human; D011971:Receptors, Immunologic; C505719:TNFSF13B protein, human; D015534:Trans-Activators
keywords: 3′UTR; Gene expression; KLRG1; eQTL; miR-584-5p; rs1805672