TY - JOUR

T1 - Mutation analysis of NR0B2 among 1545 Danish men identifies a novel c.278G>A (p.G93D) variant with reduced functional activity

AU - Echwald, Søren Morgenthaler

AU - Andersen, Kirstine L

AU - Sørensen, Thorkild I A

AU - Larsen, Lesli H

AU - Andersen, Teis

AU - Tonooka, Naoko

AU - Tomura, Hideaki

AU - Takeda, Jun

AU - Pedersen, Oluf

N1 - Copyright 2004 Wiley-Liss, Inc.

PY - 2004

Y1 - 2004

N2 - Variations of the small heterodimer partner (SHP, NR0B2) gene, an atypical nuclear receptor that inhibits transactivation by hepatocyte nuclear factor (HNF)-4alpha, are associated with obesity among Japanese. The purpose of the study was to evaluate the prevalence of SHP variants among obese Danish men. Using combined SSCP and heteroduplex analysis, we analyzed the entire coding region of SHP for variants in a cohort of 750 Danish men with early-onset obesity and genotyped a cohort of 795 nonobese control subjects using PCR-RFLP. Functional analyses of the identified coding region variants were performed in both MIN6-m9 and HepG2 cell lines. A total of five novel variants, including three missense variants (c.100C>G [p.R34G], c.278G>A [p.G93D], and c.415C>A [p.P139H]) and two silent variants (c.65C>T [p.Y22Y] and c.339G>A [p.P113P]) were identified. Moreover, the previously reported c.512G>C [p.G171A] polymorphism was identified. The 171A allele was not associated with obesity (p = 0.07). The 34G, 93D, and 139H-alleles were rare variants, which were found only among obese subjects. Among the four coding region variants, the 93D-allele showed a reduced in vitro inhibition of the HNF-4alpha transactivation of the HNF-1alpha promoter expression when expressed in MIN6-m9 and HepG2 cell lines (p

AB - Variations of the small heterodimer partner (SHP, NR0B2) gene, an atypical nuclear receptor that inhibits transactivation by hepatocyte nuclear factor (HNF)-4alpha, are associated with obesity among Japanese. The purpose of the study was to evaluate the prevalence of SHP variants among obese Danish men. Using combined SSCP and heteroduplex analysis, we analyzed the entire coding region of SHP for variants in a cohort of 750 Danish men with early-onset obesity and genotyped a cohort of 795 nonobese control subjects using PCR-RFLP. Functional analyses of the identified coding region variants were performed in both MIN6-m9 and HepG2 cell lines. A total of five novel variants, including three missense variants (c.100C>G [p.R34G], c.278G>A [p.G93D], and c.415C>A [p.P139H]) and two silent variants (c.65C>T [p.Y22Y] and c.339G>A [p.P113P]) were identified. Moreover, the previously reported c.512G>C [p.G171A] polymorphism was identified. The 171A allele was not associated with obesity (p = 0.07). The 34G, 93D, and 139H-alleles were rare variants, which were found only among obese subjects. Among the four coding region variants, the 93D-allele showed a reduced in vitro inhibition of the HNF-4alpha transactivation of the HNF-1alpha promoter expression when expressed in MIN6-m9 and HepG2 cell lines (p

KW - Adolescent

KW - Adult

KW - Age of Onset

KW - Alleles

KW - Amino Acid Sequence

KW - Case-Control Studies

KW - Cell Line, Tumor

KW - DNA Mutational Analysis

KW - Genetic Variation

KW - Humans

KW - Male

KW - Middle Aged

KW - Molecular Sequence Data

KW - Mutation, Missense

KW - Netherlands

KW - Obesity

KW - Phenotype

KW - Polymerase Chain Reaction

KW - Polymorphism, Restriction Fragment Length

KW - Polymorphism, Single-Stranded Conformational

KW - Receptors, Cytoplasmic and Nuclear

U2 - 10.1002/humu.20090

DO - 10.1002/humu.20090

M3 - Journal article

C2 - 15459958

VL - 24

SP - 381

EP - 387

JO - Human Mutation

JF - Human Mutation

SN - 1059-7794

IS - 5

ER -