title: Functional analysis of differences in transcriptional activity conferred by genetic variants in the 5' flanking region of the IL12RB2 gene.
abstract: Interleukin 12 receptor β chain (IL12RB2) is a crucial regulatory factor involved in cell-mediated immune responses, and genetic variants of the gene encoding IL12RB2 are associated with susceptibility to various immune-related diseases. We previously demonstrated that haplotypes with single nucleotide polymorphisms (SNPs) in the 5' flanking region of IL12RB2, including -1035A>G (rs3762315) and -1023A>G (rs3762316), affect the expression of IL12RB2, thereby altering susceptibility to leprosy and periodontal diseases. In the present study, we identified transcription factors associated with the haplotype-specific transcriptional activity of IL12RB2 in T cells and NK cells. The -1023G polymorphism was found to create a consensus binding site for the transcription factor activating protein (AP)-1, and enzyme-linked immunosorbent assay (ELISA)-based binding assays showed that these SNPs enhanced AP-1 binding to this region. In reporter assays, suppression of JunB expression using siRNA eliminated differences in the -1035G/-1023G and -1035A/-1023A regions containing IL12RB2 promoter activity in Jurkat T cells and NK3.3 cells. These results suggested that the -1035/-1023 polymorphisms created differential binding affinities for JunB that could lead to differential IL12RB2 expression. Moreover, the -1035G and -1035A alleles formed binding sites for GATA-3 and myocyte enhancer factor-2 (MEF-2), respectively. Our data indicated that in addition to JunB, the SNP at -1035/-1023 influenced GATA-3 and MEF-2 binding affinity, potentially altering IL12RB2 transcriptional activity. These findings confirm the effects of rs3762315 and rs3762316 on IL12RB2 transcription. These genetic variants may alter cellular activation of T cells and NK cells and modify cell-mediated immune responses.
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan. nkogoe@hyo-med.ac.jp.
authors.forename: Nahoko
authors.initials: N
authors.lastname: Kato-Kogoe
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Ohyama
authors.affiliation: Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, Matsudo, Japan.
authors.forename: Soichiro
authors.initials: S
authors.lastname: Okano
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Yamanegi
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Yamada
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Hata
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nishiura
authors.affiliation: Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo, Matsudo, Japan.
authors.forename: Yoshimitsu
authors.initials: Y
authors.lastname: Abiko
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Terada
authors.affiliation: Department of Pathology, Hyogo College of Medicine, Nishinomiya, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Nakasho
mesh_terms: D024506:5' Flanking Region; D050990:GATA3 Transcription Factor / Q000378:metabolism; D006239:Haplotypes; D006801:Humans; D019169:Jurkat Cells; D007694:Killer Cells, Natural / Q000378:metabolism; D064326:MEF2 Transcription Factors / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D053707:Receptors, Interleukin-12 / Q000235:genetics* / Q000378:metabolism*; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D050990:GATA3 Transcription Factor; C508439:IL12RB2 protein, human; D064326:MEF2 Transcription Factors; D053707:Receptors, Interleukin-12; D018808:Transcription Factor AP-1
keywords: Interleukin 12 receptor β chain; NK cells; Single nucleotide polymorphism; T cells