title: The single nucleotide variant rs12722489 determines differential estrogen receptor binding and enhancer properties of an IL2RA intronic region.
abstract: We studied functional effect of rs12722489 single nucleotide polymorphism located in the first intron of human IL2RA gene on transcriptional regulation. This polymorphism is associated with multiple autoimmune conditions (rheumatoid arthritis, multiple sclerosis, Crohn's disease, and ulcerative colitis). Analysis in silico suggested significant difference in the affinity of estrogen receptor (ER) binding site between alternative allelic variants, with stronger predicted affinity for the risk (G) allele. Electrophoretic mobility shift assay showed that purified human ERα bound only G variant of a 32-bp genomic sequence containing rs12722489. Chromatin immunoprecipitation demonstrated that endogenous human ERα interacted with rs12722489 genomic region in vivo and DNA pull-down assay confirmed differential allelic binding of amplified 189-bp genomic fragments containing rs12722489 with endogenous human ERα. In a luciferase reporter assay, a kilobase-long genomic segment containing G but not A allele of rs12722489 demonstrated enhancer properties in MT-2 cell line, an HTLV-1 transformed human cell line with a regulatory T cell phenotype.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Marina A
authors.identifier: http://orcid.org/0000-0001-5294-970X
authors.initials: MA
authors.lastname: Afanasyeva
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Lidia V
authors.initials: LV
authors.lastname: Putlyaeva
authors.affiliation: Moscow Institute of Physics and Technology, Moscow, Russia.
authors.forename: Denis E
authors.initials: DE
authors.lastname: Demin
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.|Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia.|Skolkovo Institute of Science and Technology, Skolkovo Innovation Center, Moscow, Russia.
authors.forename: Ivan V
authors.initials: IV
authors.lastname: Kulakovskiy
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Ilya E
authors.initials: IE
authors.lastname: Vorontsov
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Fridman
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.|Moscow Institute of Physics and Technology, Moscow, Russia.|Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Vsevolod J
authors.initials: VJ
authors.lastname: Makeev
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.|Moscow Institute of Physics and Technology, Moscow, Russia.|Faculty of Biology, Lomonosov Moscow State University, Moscow, Russia.
authors.forename: Dmitry V
authors.initials: DV
authors.lastname: Kuprash
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Anton M
authors.initials: AM
authors.lastname: Schwartz
mesh_terms: D000483:Alleles; D001483:Base Sequence; D001665:Binding Sites; D002461:Cell Line, Transformed; D047369:Chromatin Immunoprecipitation; D024202:Electrophoretic Mobility Shift Assay; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D015368:Human T-lymphotropic virus 1 / Q000235:genetics; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit / Q000235:genetics* / Q000378:metabolism; D007438:Introns; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D011485:Protein Binding; D020218:Response Elements; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000378:metabolism*
chemical_list: D047628:Estrogen Receptor alpha; C508017:IL2RA protein, human; D053645:Interleukin-2 Receptor alpha Subunit; D008156:Luciferases