Establishment of a simple diagnostic method for the detection of mutations of insulin receptor gene in Non-Insulin Dependent Diabetes Mellitus

• Project/Area Number: 02557017
• Research Category: Grant-in-Aid for Developmental Scientific Research (B)
• Allocation Type: Single-year Grants
• Research Field: Pathological medical chemistry
• Research Institution: The University of Tokushima
• Principal Investigator: EBINA Yousuke The University of Tokushima, Institute for Enzyme Research, Professor, 酵素科学研究センター, 教授 (00112227)
• Co-Investigator(Kenkyū-buntansha): MATSUOKA Yukio Chyugai pharmaceutical Co. Ltd., 応用研究所, 主任 ; HAYASHI Hideki The University of Tokushima Institute for Enzyme Research, Research Associate, 酵素科学研究センター, 助手 (10218589) ; MURAKAMI Takashi The University of Tokushima Institute for Enzyme Research, Research Associate, 酵素科学研究センター, 助手 (40210009)
• Project Period (FY): 1990 – 1991
• Project Status: Completed (Fiscal Year 1991)
• Budget Amount: ¥15,400,000 (Direct Cost: ¥15,400,000); Fiscal Year 1991: ¥4,800,000 (Direct Cost: ¥4,800,000); Fiscal Year 1990: ¥10,600,000 (Direct Cost: ¥10,600,000)
• Keywords: NIDDM / Insulin Receptor / Mutation

Research Abstract

Our previous reports (Science (1989) 24S, 63-66 ; The Lancet (1990) 335, 1179-1181) suggested the possibility that the mutations of insulin receptor gene might be involved in genetic factors of Non-Insulin Dependent Diabetes Mellitus (NIDDM). Therefore, we started to examine the frequency of the abnormalities of insulin receptor gene in NIDDM. We purified the genomic DNAs from typical 50 NIDDM patients. Eleven exons of beta subunit from 50 patients were amplified and the nucleotide sequences were determined. We found several polymorphisms in- Exon 13 and 17. There was the mutation of amino acid sequences at the same point in three NIDDM patients. We are going to check the function of the receptor which has the amino acid substitution using the expression system and the artificial mutation on the CDNA. We arealso going to examine the mutations in the 11 exons of alpha subunit. Our final goal is to establish a simple diagn ostic method for the detection of mutations of insulin receptor gene in NIDDM.

Research Products

• [Publications] Murakami,T.,Ebina,Y.et al.: "Identification of Two Enhancer Elements in the Gene Encoding the Type 1 Glucose Transporter from the Mouse Which are Responsive to Serum,Growth Factor and Oncogenes." j.Biol.Chem.
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Hayashi,H.,Ebina,Y.et al.: "Phosphorylation in vitro of the 85 kDa subunit of phosphatidylinositol 3-kinase and its possible activation by insulin receptor tyrosine kinase." Biochem.J.280. 769-775 (1991)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Araki,E.,Ebina,Y.et al.: "A cluster of four Sp1 binding sites required for efficient expression of the human insulin receptor gene." J.Biol.Chem.266. 3944-3948 (1991)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Hayashi,H.,Ebina,Y.et al.: "Insulin treatment stimulates the tyrosine phosphorylation of the 85-kDa subunit of phosphatidylinositol 3-kinase in vivo and possibly stimulates its specific activity." J.Biol.Chem.
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Hayashi,H.,Ebina,Y.et al.: "The 85-kDa subunit of phosphatidylinositol 3-kinase is phosphorylated at tyrosine 607 by insulin receptor in vitro and possibly in vivo." J.Biol.Chem.
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Murakami, T., Ebina, Y. et al.: "Identificaton of Two Enhancer Elements in the Gene Encoding the Type 1 Glucose Transporter from the Mouse Which are Responsive to Serum, Growth Factor and Oncogenes." J. Biol. Chem.
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hayashi, H., Ebina, Y. et al.: "Phosphorylation in vitro of the 85 kDa subunit of phosphatidylinoisitol 3-kinase and its possible activation by insulin receptor tyrosine kinse." Biochem. J.280. 769-775 (1991)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Araki, E., Ebina, Y. et al.: "A cluster of four SP1 binding sites required for efficient expression of the human insulin receptor gene." J. Biol. Chem. 266. 3944-3948 (1991)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hayashi, H., Ebina, Y. et al.: "Insulin treatemnt stimulates the tyrosine phosphorylation of the 85-kDa subunit of phosphatidylinositol 3-kinase in vivo and possibly stimulates its specific activity" J. Biol. Chem.
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Hayashi, H., Ebina, Y. et al.: "The 85-kDa subunit of phosphatidylinositol 3-kinase is phosphorylated at tyrosine 607 by insulin receptor in vitro and possibly in vivo." J. Biol. Chem.
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Murakami,Y.,Ebina,Y.et al.: "Inentification of Two Enhancer Elements in the Gene Encoding the Type 1 Glucose Transporter from the Mouse Which are Responsive to Serum,Growth Factor and Oncogenes." J.Biol.Chem.
• [Publications] Hayashi,H.,Ebina,Y.et al.: "Phosphprylation in vitro of the 85 kDa subunit of phosphatidylinositol 3-Kinase and its Possible activation by insulin receptor tyrosine Kinase." Biochem.J.280. 769-775 (1991)
• [Publications] E.Araki,E.,Ebina,Y.et al.: "A cluster of four Spl binding sites required for efficient expression of the human insulin receptor gene." J.Biol.Chem.266. 3944-3948 (1991)
• [Publications] Shimada,F.,Ebina,Y.et al.: "Insulinーresistant diabetes associated with partial deletion of insulinーreceptor gene." The Lancet. 335. 1179-1181 (1990)
• [Publications] Araki,E.,Edina,Y.et al.: "A cluster of four Spl binding sites required for efficient expression of the human insulin receptor gene." J.Biol.Chem.
• [Publications] Hayashi,H.,Ebina,Y.et al.: "Evidence that phosphatidylinositol 3 Kinase is a substrate for insulin receptor tyrosine kinase." J.Biol.Chem.