RRC ID 41654
著者 Iwamoto K, Mori H, Okazawa H, Hashiramoto M, Kasuga M.
タイトル Identification of a single nucleotide polymorphism showing no insulin-mediated suppression of the promoter activity in the human insulin receptor substrate 2 gene.
ジャーナル Diabetologia
Abstract AIMS/HYPOTHESIS:To understand the transcriptional regulation and to investigate the pathological influence upon Type II (non-insulin-dependent) diabetes mellitus of insulin receptor substrate 2 ( IRS2), the 5' flanking region of the human IRS2 gene was cloned and screened in Japanese diabetic patients.
METHODS:Luciferase reporter assay and electrophoretic mobility shift assay (EMSA) were combined in HepG2, Fao, RINm5F, and HeLa cells to characterise the human IRS2 promoter region. Single nucleotide polymorphisms (SNPs) were identified in Japanese Type II diabetic patients by sequencing and were genotyped.
RESULTS:The proximal 2399 bp of the 5' flanking region of the human IRS2 gene was cloned. A core promoter region was extended between nucleotide positions -834 and -557 (relative to the translation initiation site). The region [(-758)AGGGGGAGGG(-749)] that appears important in the positive regulation of IRS2transcription was identified by EMSA with (32)P-labelled double-stranded oligonucleotides encompassing regions protected from DNase I digestion by nuclear extract of HepG2 cells. Two SNPs (-765 C/ T and -2062 T/ C), identified by screening Japanese Type II diabetic patients, were not associated with Type II diabetes. IRS2-driven reporter activity in the plasmid containing thymine at -765 was not suppressed by insulin when measured in Fao cells.
CONCLUSION/INTERPRETATION:The 5' flanking sequence of the human IRS2 was investigated and two SNPs were identified. The SNP at -765 was suggested to be involved in the insulin-mediated regulation of the transcriptional activity of IRS2.
巻・号 45(8)
ページ 1182-95
公開日 2002-8-1
DOI 10.1007/s00125-002-0871-0
PMID 12189449
MeSH 5' Untranslated Regions / genetics Aged Base Sequence / genetics Cells, Cultured DNA Footprinting Deoxyribonuclease I / genetics Diabetes Mellitus, Type 2 / genetics* Diabetes Mellitus, Type 2 / metabolism* Electrophoresis Female Humans Hypoglycemic Agents / metabolism* Insulin / metabolism* Insulin Receptor Substrate Proteins Intracellular Signaling Peptides and Proteins Male Middle Aged Molecular Sequence Data Mutation / physiology Phosphoproteins / genetics* Polymorphism, Single Nucleotide* Promoter Regions, Genetic / physiology*
IF 7.518
引用数 6
WOS 分野 ENDOCRINOLOGY & METABOLISM
リソース情報
ヒト・動物細胞