Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1997 Dec 15;100(12):3164–3172. doi: 10.1172/JCI119872

Differential regulation of insulin receptor substrates-1 and -2 (IRS-1 and IRS-2) and phosphatidylinositol 3-kinase isoforms in liver and muscle of the obese diabetic (ob/ob) mouse.

N J Kerouz 1, D Hörsch 1, S Pons 1, C R Kahn 1
PMCID: PMC508530  PMID: 9399964

Abstract

Intracellular insulin signaling involves a series of alternative and complementary pathways created by the multiple substrates of the insulin receptor (IRS) and the various isoforms of SH2 domain signaling molecules that can interact with these substrates. In this study, we have evaluated the roles of IRS-1 and IRS-2 in signaling to the phosphatidylinositol (PI) 3-kinase pathway in the ob/ob mouse, a model of the insulin resistance of obesity and non-insulin-dependent diabetes mellitus. We find that the levels of expression of both IRS-1 and IRS-2 are decreased approximately 50% in muscle, whereas in liver the decrease is significantly greater for IRS-2 (72%) than for IRS-1 (29%). This results in differential decreases in IRS-1 and IRS-2 phosphorylation, docking of the p85alpha regulatory subunit of PI 3-kinase, and activation of this enzyme in these two insulin target tissues. In ob/ob liver there is also a change in expression of the alternatively spliced isoforms of the regulatory subunits for PI 3-kinase that was detected at the protein and mRNA level. This resulted in a 45% decrease in the p85alpha form of PI 3-kinase, a ninefold increase in the AS53/p55alpha, and a twofold increase in p50alpha isoforms. Thus, there are multiple alterations in the early steps of insulin signaling in the ob/ob mouse, with differential regulation of IRS-1 and IRS-2, various PI 3-kinase regulatory isoforms, and a lack of compensation for the decrease in insulin signaling by any of the known alternative pathways at these levels.

Full Text

The Full Text of this article is available as a PDF (523.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Antonetti D. A., Algenstaedt P., Kahn C. R. Insulin receptor substrate 1 binds two novel splice variants of the regulatory subunit of phosphatidylinositol 3-kinase in muscle and brain. Mol Cell Biol. 1996 May;16(5):2195–2203. doi: 10.1128/mcb.16.5.2195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Araki E., Lipes M. A., Patti M. E., Brüning J. C., Haag B., 3rd, Johnson R. S., Kahn C. R. Alternative pathway of insulin signalling in mice with targeted disruption of the IRS-1 gene. Nature. 1994 Nov 10;372(6502):186–190. doi: 10.1038/372186a0. [DOI] [PubMed] [Google Scholar]
  3. Backer J. M., Myers M. G., Jr, Shoelson S. E., Chin D. J., Sun X. J., Miralpeix M., Hu P., Margolis B., Skolnik E. Y., Schlessinger J. Phosphatidylinositol 3'-kinase is activated by association with IRS-1 during insulin stimulation. EMBO J. 1992 Sep;11(9):3469–3479. doi: 10.1002/j.1460-2075.1992.tb05426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brüning J. C., Winnay J., Bonner-Weir S., Taylor S. I., Accili D., Kahn C. R. Development of a novel polygenic model of NIDDM in mice heterozygous for IR and IRS-1 null alleles. Cell. 1997 Feb 21;88(4):561–572. doi: 10.1016/s0092-8674(00)81896-6. [DOI] [PubMed] [Google Scholar]
  5. Cheatham B., Kahn C. R. Insulin action and the insulin signaling network. Endocr Rev. 1995 Apr;16(2):117–142. doi: 10.1210/edrv-16-2-117. [DOI] [PubMed] [Google Scholar]
  6. Cheatham B., Vlahos C. J., Cheatham L., Wang L., Blenis J., Kahn C. R. Phosphatidylinositol 3-kinase activation is required for insulin stimulation of pp70 S6 kinase, DNA synthesis, and glucose transporter translocation. Mol Cell Biol. 1994 Jul;14(7):4902–4911. doi: 10.1128/mcb.14.7.4902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chua S. C., Jr, Chung W. K., Wu-Peng X. S., Zhang Y., Liu S. M., Tartaglia L., Leibel R. L. Phenotypes of mouse diabetes and rat fatty due to mutations in the OB (leptin) receptor. Science. 1996 Feb 16;271(5251):994–996. doi: 10.1126/science.271.5251.994. [DOI] [PubMed] [Google Scholar]
  8. Endemann G., Yonezawa K., Roth R. A. Phosphatidylinositol kinase or an associated protein is a substrate for the insulin receptor tyrosine kinase. J Biol Chem. 1990 Jan 5;265(1):396–400. [PubMed] [Google Scholar]
  9. Escobedo J. A., Navankasattusas S., Kavanaugh W. M., Milfay D., Fried V. A., Williams L. T. cDNA cloning of a novel 85 kd protein that has SH2 domains and regulates binding of PI3-kinase to the PDGF beta-receptor. Cell. 1991 Apr 5;65(1):75–82. doi: 10.1016/0092-8674(91)90409-r. [DOI] [PubMed] [Google Scholar]
  10. Folli F., Saad M. J., Backer J. M., Kahn C. R. Regulation of phosphatidylinositol 3-kinase activity in liver and muscle of animal models of insulin-resistant and insulin-deficient diabetes mellitus. J Clin Invest. 1993 Oct;92(4):1787–1794. doi: 10.1172/JCI116768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Freeman R. M., Jr, Plutzky J., Neel B. G. Identification of a human src homology 2-containing protein-tyrosine-phosphatase: a putative homolog of Drosophila corkscrew. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11239–11243. doi: 10.1073/pnas.89.23.11239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fruman D. A., Cantley L. C., Carpenter C. L. Structural organization and alternative splicing of the murine phosphoinositide 3-kinase p85 alpha gene. Genomics. 1996 Oct 1;37(1):113–121. doi: 10.1006/geno.1996.0527. [DOI] [PubMed] [Google Scholar]
  13. Hadari Y. R., Tzahar E., Nadiv O., Rothenberg P., Roberts C. T., Jr, LeRoith D., Yarden Y., Zick Y. Insulin and insulinomimetic agents induce activation of phosphatidylinositol 3'-kinase upon its association with pp185 (IRS-1) in intact rat livers. J Biol Chem. 1992 Sep 5;267(25):17483–17486. [PubMed] [Google Scholar]
  14. Heydrick S. J., Jullien D., Gautier N., Tanti J. F., Giorgetti S., Van Obberghen E., Le Marchand-Brustel Y. Defect in skeletal muscle phosphatidylinositol-3-kinase in obese insulin-resistant mice. J Clin Invest. 1993 Apr;91(4):1358–1366. doi: 10.1172/JCI116337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hiles I. D., Otsu M., Volinia S., Fry M. J., Gout I., Dhand R., Panayotou G., Ruiz-Larrea F., Thompson A., Totty N. F. Phosphatidylinositol 3-kinase: structure and expression of the 110 kd catalytic subunit. Cell. 1992 Aug 7;70(3):419–429. doi: 10.1016/0092-8674(92)90166-a. [DOI] [PubMed] [Google Scholar]
  16. Holgado-Madruga M., Emlet D. R., Moscatello D. K., Godwin A. K., Wong A. J. A Grb2-associated docking protein in EGF- and insulin-receptor signalling. Nature. 1996 Feb 8;379(6565):560–564. doi: 10.1038/379560a0. [DOI] [PubMed] [Google Scholar]
  17. Inukai K., Anai M., Van Breda E., Hosaka T., Katagiri H., Funaki M., Fukushima Y., Ogihara T., Yazaki Y., Kikuchi A novel 55-kDa regulatory subunit for phosphatidylinositol 3-kinase structurally similar to p55PIK Is generated by alternative splicing of the p85alpha gene. J Biol Chem. 1996 Mar 8;271(10):5317–5320. doi: 10.1074/jbc.271.10.5317. [DOI] [PubMed] [Google Scholar]
  18. Inukai K., Funaki M., Ogihara T., Katagiri H., Kanda A., Anai M., Fukushima Y., Hosaka T., Suzuki M., Shin B. C. p85alpha gene generates three isoforms of regulatory subunit for phosphatidylinositol 3-kinase (PI 3-Kinase), p50alpha, p55alpha, and p85alpha, with different PI 3-kinase activity elevating responses to insulin. J Biol Chem. 1997 Mar 21;272(12):7873–7882. doi: 10.1074/jbc.272.12.7873. [DOI] [PubMed] [Google Scholar]
  19. Kahn C. R., Neville D. M., Jr, Roth J. Insulin-receptor interaction in the obese-hyperglycemic mouse. A model of insulin resistance. J Biol Chem. 1973 Jan 10;248(1):244–250. [PubMed] [Google Scholar]
  20. Kaliman P., Viñals F., Testar X., Palacín M., Zorzano A. Phosphatidylinositol 3-kinase inhibitors block differentiation of skeletal muscle cells. J Biol Chem. 1996 Aug 9;271(32):19146–19151. doi: 10.1074/jbc.271.32.19146. [DOI] [PubMed] [Google Scholar]
  21. Kapeller R., Cantley L. C. Phosphatidylinositol 3-kinase. Bioessays. 1994 Aug;16(8):565–576. doi: 10.1002/bies.950160810. [DOI] [PubMed] [Google Scholar]
  22. Kellerer M., Sesti G., Seffer E., Obermaier-Kusser B., Pongratz D. E., Mosthaf L., Häring H. U. Altered pattern of insulin receptor isotypes in skeletal muscle membranes of type 2 (non-insulin-dependent) diabetic subjects. Diabetologia. 1993 Jul;36(7):628–632. doi: 10.1007/BF00404072. [DOI] [PubMed] [Google Scholar]
  23. King P. A., Horton E. D., Hirshman M. F., Horton E. S. Insulin resistance in obese Zucker rat (fa/fa) skeletal muscle is associated with a failure of glucose transporter translocation. J Clin Invest. 1992 Oct;90(4):1568–1575. doi: 10.1172/JCI116025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klippel A., Escobedo J. A., Hirano M., Williams L. T. The interaction of small domains between the subunits of phosphatidylinositol 3-kinase determines enzyme activity. Mol Cell Biol. 1994 Apr;14(4):2675–2685. doi: 10.1128/mcb.14.4.2675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lavan B. E., Fantin V. R., Chang E. T., Lane W. S., Keller S. R., Lienhard G. E. A novel 160-kDa phosphotyrosine protein in insulin-treated embryonic kidney cells is a new member of the insulin receptor substrate family. J Biol Chem. 1997 Aug 22;272(34):21403–21407. doi: 10.1074/jbc.272.34.21403. [DOI] [PubMed] [Google Scholar]
  26. Lavan B. E., Lienhard G. E. The insulin-elicited 60-kDa phosphotyrosine protein in rat adipocytes is associated with phosphatidylinositol 3-kinase. J Biol Chem. 1993 Mar 15;268(8):5921–5928. [PubMed] [Google Scholar]
  27. Mosthaf L., Vogt B., Häring H. U., Ullrich A. Altered expression of insulin receptor types A and B in the skeletal muscle of non-insulin-dependent diabetes mellitus patients. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4728–4730. doi: 10.1073/pnas.88.11.4728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Myers M. G., Jr, Sun X. J., Cheatham B., Jachna B. R., Glasheen E. M., Backer J. M., White M. F. IRS-1 is a common element in insulin and insulin-like growth factor-I signaling to the phosphatidylinositol 3'-kinase. Endocrinology. 1993 Apr;132(4):1421–1430. doi: 10.1210/endo.132.4.8384986. [DOI] [PubMed] [Google Scholar]
  29. Norgren S., Zierath J., Wedell A., Wallberg-Henriksson H., Luthman H. Regulation of human insulin receptor RNA splicing in vivo. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1465–1469. doi: 10.1073/pnas.91.4.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Otsu M., Hiles I., Gout I., Fry M. J., Ruiz-Larrea F., Panayotou G., Thompson A., Dhand R., Hsuan J., Totty N. Characterization of two 85 kd proteins that associate with receptor tyrosine kinases, middle-T/pp60c-src complexes, and PI3-kinase. Cell. 1991 Apr 5;65(1):91–104. doi: 10.1016/0092-8674(91)90411-q. [DOI] [PubMed] [Google Scholar]
  31. Patti M. E., Sun X. J., Bruening J. C., Araki E., Lipes M. A., White M. F., Kahn C. R. 4PS/insulin receptor substrate (IRS)-2 is the alternative substrate of the insulin receptor in IRS-1-deficient mice. J Biol Chem. 1995 Oct 20;270(42):24670–24673. doi: 10.1074/jbc.270.42.24670. [DOI] [PubMed] [Google Scholar]
  32. Pons S., Asano T., Glasheen E., Miralpeix M., Zhang Y., Fisher T. L., Myers M. G., Jr, Sun X. J., White M. F. The structure and function of p55PIK reveal a new regulatory subunit for phosphatidylinositol 3-kinase. Mol Cell Biol. 1995 Aug;15(8):4453–4465. doi: 10.1128/mcb.15.8.4453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Saad M. J., Araki E., Miralpeix M., Rothenberg P. L., White M. F., Kahn C. R. Regulation of insulin receptor substrate-1 in liver and muscle of animal models of insulin resistance. J Clin Invest. 1992 Nov;90(5):1839–1849. doi: 10.1172/JCI116060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Saad M. J., Folli F., Kahn J. A., Kahn C. R. Modulation of insulin receptor, insulin receptor substrate-1, and phosphatidylinositol 3-kinase in liver and muscle of dexamethasone-treated rats. J Clin Invest. 1993 Oct;92(4):2065–2072. doi: 10.1172/JCI116803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Skolnik E. Y., Lee C. H., Batzer A., Vicentini L. M., Zhou M., Daly R., Myers M. J., Jr, Backer J. M., Ullrich A., White M. F. The SH2/SH3 domain-containing protein GRB2 interacts with tyrosine-phosphorylated IRS1 and Shc: implications for insulin control of ras signalling. EMBO J. 1993 May;12(5):1929–1936. doi: 10.1002/j.1460-2075.1993.tb05842.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Skolnik E. Y., Margolis B., Mohammadi M., Lowenstein E., Fischer R., Drepps A., Ullrich A., Schlessinger J. Cloning of PI3 kinase-associated p85 utilizing a novel method for expression/cloning of target proteins for receptor tyrosine kinases. Cell. 1991 Apr 5;65(1):83–90. doi: 10.1016/0092-8674(91)90410-z. [DOI] [PubMed] [Google Scholar]
  37. Skouteris G. G., Georgakopoulos E. Hepatocyte growth factor-induced proliferation of primary hepatocytes is mediated by activation of phosphatidylinositol 3-kinase. Biochem Biophys Res Commun. 1996 Jan 5;218(1):229–233. doi: 10.1006/bbrc.1996.0040. [DOI] [PubMed] [Google Scholar]
  38. Smith-Hall J., Pons S., Patti M. E., Burks D. J., Yenush L., Sun X. J., Kahn C. R., White M. F. The 60 kDa insulin receptor substrate functions like an IRS protein (pp60IRS3) in adipose cells. Biochemistry. 1997 Jul 8;36(27):8304–8310. doi: 10.1021/bi9630974. [DOI] [PubMed] [Google Scholar]
  39. Soli A. H., Kahn C. R., Neville D. M., Jr, Roth J. Insulin receptor deficiency in genetic and acquired obesity. J Clin Invest. 1975 Oct;56(4):769–780. doi: 10.1172/JCI108155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sugimoto S., Lechleider R. J., Shoelson S. E., Neel B. G., Walsh C. T. Expression, purification, and characterization of SH2-containing protein tyrosine phosphatase, SH-PTP2. J Biol Chem. 1993 Oct 25;268(30):22771–22776. [PubMed] [Google Scholar]
  41. Sun X. J., Crimmins D. L., Myers M. G., Jr, Miralpeix M., White M. F. Pleiotropic insulin signals are engaged by multisite phosphorylation of IRS-1. Mol Cell Biol. 1993 Dec;13(12):7418–7428. doi: 10.1128/mcb.13.12.7418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sun X. J., Rothenberg P., Kahn C. R., Backer J. M., Araki E., Wilden P. A., Cahill D. A., Goldstein B. J., White M. F. Structure of the insulin receptor substrate IRS-1 defines a unique signal transduction protein. Nature. 1991 Jul 4;352(6330):73–77. doi: 10.1038/352073a0. [DOI] [PubMed] [Google Scholar]
  43. Sun X. J., Wang L. M., Zhang Y., Yenush L., Myers M. G., Jr, Glasheen E., Lane W. S., Pierce J. H., White M. F. Role of IRS-2 in insulin and cytokine signalling. Nature. 1995 Sep 14;377(6545):173–177. doi: 10.1038/377173a0. [DOI] [PubMed] [Google Scholar]
  44. Tamemoto H., Kadowaki T., Tobe K., Yagi T., Sakura H., Hayakawa T., Terauchi Y., Ueki K., Kaburagi Y., Satoh S. Insulin resistance and growth retardation in mice lacking insulin receptor substrate-1. Nature. 1994 Nov 10;372(6502):182–186. doi: 10.1038/372182a0. [DOI] [PubMed] [Google Scholar]
  45. Tobe K., Tamemoto H., Yamauchi T., Aizawa S., Yazaki Y., Kadowaki T. Identification of a 190-kDa protein as a novel substrate for the insulin receptor kinase functionally similar to insulin receptor substrate-1. J Biol Chem. 1995 Mar 17;270(11):5698–5701. doi: 10.1074/jbc.270.11.5698. [DOI] [PubMed] [Google Scholar]
  46. Tomiyama K., Nakata H., Sasa H., Arimura S., Nishio E., Watanabe Y. Wortmannin, a specific phosphatidylinositol 3-kinase inhibitor, inhibits adipocytic differentiation of 3T3-L1 cells. Biochem Biophys Res Commun. 1995 Jul 6;212(1):263–269. doi: 10.1006/bbrc.1995.1965. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES