PMID:8513971

Citation	Araki, E, Sun, XJ, Haag, BL 3rd, Chuang, LM, Zhang, Y, Yang-Feng, TL, White, MF and Kahn, CR (1993) Human skeletal muscle insulin receptor substrate-1. Characterization of the cDNA, gene, and chromosomal localization. Diabetes 42:1041-54
Abstract	Insulin receptor substrate-1 is a major substrate of insulin receptor Tyr kinase. We have now cloned the IRS-1 cDNA from human skeletal muscle, one of the most important target tissues of insulin action, localized and cloned the human IRS-1 gene, and studied the expression of the protein in Chinese hamster ovary cells. Human IRS-1 cDNA encodes a 1242 amino acid sequence that is 88% identical with rat liver IRS-1. The 14 potential Tyr phosphorylation sites include 6 Tyr-Met-X-Met motifs and 3 Tyr-X-X-Met motifs that are completely conserved in human IRS-1. Human IRS-1 has > 50 possible Ser/Thr phosphorylation sites and one potential ATP-binding site close to the NH2-terminal. The human IRS-1 gene contains the entire 5'-untranslated region and protein coding region in a single exon and was localized on chromosome 2 q36-37 by in situ hybridization. By Northern blot analysis, IRS-1 mRNA is rare and consists of two species of 6.9 and 6 kilobase. By using quantitative polymerase chain reaction after reverse transcription of total RNA from human fetal tissues, IRS-1 mRNA could be identified in all tissues. When human IRS-1 cDNA was expressed in Chinese hamster ovary cells, the protein migrated between 170,000-180,000 M(r) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was rapidly Tyr phosphorylated upon insulin stimulation. Thus, IRS-1 is widely expressed and highly conserved across species and tissues.(ABSTRACT TRUNCATED AT 250 WORDS)
Links	PubMed
Keywords	Amino Acid Sequence; Animals; Base Sequence; CHO Cells; Chromosome Mapping; Chromosomes, Human, Pair 2; Cloning, Molecular; Cricetinae; DNA; Fetus; Gene Expression; Genomic Library; Humans; In Situ Hybridization; Insulin Receptor Substrate Proteins; Liver/metabolism; Molecular Sequence Data; Muscles/metabolism; Oligodeoxyribonucleotides; Phosphoproteins/biosynthesis; Phosphoproteins/genetics; Polymerase Chain Reaction; RNA, Messenger/biosynthesis; RNA, Messenger/metabolism; Rats; Sequence Homology, Amino Acid; Transcription, Genetic; Transfection
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