Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Aug 6;93(16):8401–8406. doi: 10.1073/pnas.93.16.8401

Evidence for an insulin receptor substrate 1 independent insulin signaling pathway that mediates insulin-responsive glucose transporter (GLUT4) translocation.

A J Morris 1, S S Martin 1, T Haruta 1, J G Nelson 1, P Vollenweider 1, T A Gustafson 1, M Mueckler 1, D W Rose 1, J M Olefsky 1
PMCID: PMC38683  PMID: 8710883

Abstract

Interaction of the activated insulin receptor (IR) with its substrate, insulin receptor substrate 1 (IRS-1), via the phosphotyrosine binding domain of IRS-1 and the NPXY motif centered at phosphotyrosine 960 of the IR, is important for IRS-1 phosphorylation. We investigated the role of this interaction in the insulin signaling pathway that stimulates glucose transport. Utilizing microinjection of competitive inhibitory reagents in 3T3-L1 adipocytes, we have found that disruption of the IR/IRS-1 interaction has no effect upon translocation of the insulin-responsive glucose transporter (GLUT4). The activity of these reagents was demonstrated by their ability to block insulin stimulation of two distinct insulin bioeffects, membrane ruffling and mitogenesis, in 3T3-L1 adipocytes and insulin-responsive rat 1 fibroblasts. These data suggest that phosphorylated IRS-1 is not an essential component of the metabolic insulin signaling pathway that leads to GLUT4 translocation, yet it appears to be required for other insulin bioeffects.

Full text

PDF
8401

Images in this article

8403Fig. 1
on p.8403
8404Fig. 2
on p.8404
8405Fig. 3
on p.8405

Selected References

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

  1. Araki E., Haag B. L., 3rd, Kahn C. R. Cloning of the mouse insulin receptor substrate-1 (IRS-1) gene and complete sequence of mouse IRS-1. Biochim Biophys Acta. 1994 Apr 28;1221(3):353–356. doi: 10.1016/0167-4889(94)90261-5. [DOI] [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. Araki E., Sun X. J., Haag B. L., 3rd, Chuang L. M., Zhang Y., Yang-Feng T. L., White M. F., Kahn C. R. Human skeletal muscle insulin receptor substrate-1. Characterization of the cDNA, gene, and chromosomal localization. Diabetes. 1993 Jul;42(7):1041–1054. doi: 10.2337/diab.42.7.1041. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Clarke J. F., Young P. W., Yonezawa K., Kasuga M., Holman G. D. Inhibition of the translocation of GLUT1 and GLUT4 in 3T3-L1 cells by the phosphatidylinositol 3-kinase inhibitor, wortmannin. Biochem J. 1994 Jun 15;300(Pt 3):631–635. doi: 10.1042/bj3000631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Craparo A., O'Neill T. J., Gustafson T. A. Non-SH2 domains within insulin receptor substrate-1 and SHC mediate their phosphotyrosine-dependent interaction with the NPEY motif of the insulin-like growth factor I receptor. J Biol Chem. 1995 Jun 30;270(26):15639–15643. doi: 10.1074/jbc.270.26.15639. [DOI] [PubMed] [Google Scholar]
  8. Gustafson T. A., He W., Craparo A., Schaub C. D., O'Neill T. J. Phosphotyrosine-dependent interaction of SHC and insulin receptor substrate 1 with the NPEY motif of the insulin receptor via a novel non-SH2 domain. Mol Cell Biol. 1995 May;15(5):2500–2508. doi: 10.1128/mcb.15.5.2500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Haney P. M., Slot J. W., Piper R. C., James D. E., Mueckler M. Intracellular targeting of the insulin-regulatable glucose transporter (GLUT4) is isoform specific and independent of cell type. J Cell Biol. 1991 Aug;114(4):689–699. doi: 10.1083/jcb.114.4.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hara K., Yonezawa K., Sakaue H., Ando A., Kotani K., Kitamura T., Kitamura Y., Ueda H., Stephens L., Jackson T. R. 1-Phosphatidylinositol 3-kinase activity is required for insulin-stimulated glucose transport but not for RAS activation in CHO cells. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7415–7419. doi: 10.1073/pnas.91.16.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Haruta T., Morris A. J., Rose D. W., Nelson J. G., Mueckler M., Olefsky J. M. Insulin-stimulated GLUT4 translocation is mediated by a divergent intracellular signaling pathway. J Biol Chem. 1995 Nov 24;270(47):27991–27994. doi: 10.1074/jbc.270.47.27991. [DOI] [PubMed] [Google Scholar]
  12. He W., O'Neill T. J., Gustafson T. A. Distinct modes of interaction of SHC and insulin receptor substrate-1 with the insulin receptor NPEY region via non-SH2 domains. J Biol Chem. 1995 Oct 6;270(40):23258–23262. doi: 10.1074/jbc.270.40.23258. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. James D. E., Brown R., Navarro J., Pilch P. F. Insulin-regulatable tissues express a unique insulin-sensitive glucose transport protein. Nature. 1988 May 12;333(6169):183–185. doi: 10.1038/333183a0. [DOI] [PubMed] [Google Scholar]
  15. Jhun B. H., Rose D. W., Seely B. L., Rameh L., Cantley L., Saltiel A. R., Olefsky J. M. Microinjection of the SH2 domain of the 85-kilodalton subunit of phosphatidylinositol 3-kinase inhibits insulin-induced DNA synthesis and c-fos expression. Mol Cell Biol. 1994 Nov;14(11):7466–7475. doi: 10.1128/mcb.14.11.7466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Keller S. R., Aebersold R., Garner C. W., Lienhard G. E. The insulin-elicited 160 kDa phosphotyrosine protein in mouse adipocytes is an insulin receptor substrate 1: identification by cloning. Biochim Biophys Acta. 1993 Mar 20;1172(3):323–326. doi: 10.1016/0167-4781(93)90222-y. [DOI] [PubMed] [Google Scholar]
  17. Liu X. J., Sorisky A., Zhu L., Pawson T. Molecular cloning of an amphibian insulin receptor substrate 1-like cDNA and involvement of phosphatidylinositol 3-kinase in insulin-induced Xenopus oocyte maturation. Mol Cell Biol. 1995 Jul;15(7):3563–3570. doi: 10.1128/mcb.15.7.3563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. O'Neill T. J., Craparo A., Gustafson T. A. Characterization of an interaction between insulin receptor substrate 1 and the insulin receptor by using the two-hybrid system. Mol Cell Biol. 1994 Oct;14(10):6433–6442. doi: 10.1128/mcb.14.10.6433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Okada T., Kawano Y., Sakakibara T., Hazeki O., Ui M. Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adipocytes. Studies with a selective inhibitor wortmannin. J Biol Chem. 1994 Feb 4;269(5):3568–3573. [PubMed] [Google Scholar]
  20. 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]
  21. Quon M. J., Butte A. J., Zarnowski M. J., Sesti G., Cushman S. W., Taylor S. I. Insulin receptor substrate 1 mediates the stimulatory effect of insulin on GLUT4 translocation in transfected rat adipose cells. J Biol Chem. 1994 Nov 11;269(45):27920–27924. [PubMed] [Google Scholar]
  22. Rose D. W., Saltiel A. R., Majumdar M., Decker S. J., Olefsky J. M. Insulin receptor substrate 1 is required for insulin-mediated mitogenic signal transduction. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):797–801. doi: 10.1073/pnas.91.2.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Staubs P. A., Reichart D. R., Saltiel A. R., Milarski K. L., Maegawa H., Berhanu P., Olefsky J. M., Seely B. L. Localization of the insulin receptor binding sites for the SH2 domain proteins p85, Syp, and GAP. J Biol Chem. 1994 Nov 4;269(44):27186–27192. [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. 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]
  27. 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]
  28. Van Horn D. J., Myers M. G., Jr, Backer J. M. Direct activation of the phosphatidylinositol 3'-kinase by the insulin receptor. J Biol Chem. 1994 Jan 7;269(1):29–32. [PubMed] [Google Scholar]
  29. White M. F., Livingston J. N., Backer J. M., Lauris V., Dull T. J., Ullrich A., Kahn C. R. Mutation of the insulin receptor at tyrosine 960 inhibits signal transmission but does not affect its tyrosine kinase activity. Cell. 1988 Aug 26;54(5):641–649. doi: 10.1016/s0092-8674(88)80008-4. [DOI] [PubMed] [Google Scholar]
  30. White M. F., Maron R., Kahn C. R. Insulin rapidly stimulates tyrosine phosphorylation of a Mr-185,000 protein in intact cells. Nature. 1985 Nov 14;318(6042):183–186. doi: 10.1038/318183a0. [DOI] [PubMed] [Google Scholar]
  31. Wolf G., Trüb T., Ottinger E., Groninga L., Lynch A., White M. F., Miyazaki M., Lee J., Shoelson S. E. PTB domains of IRS-1 and Shc have distinct but overlapping binding specificities. J Biol Chem. 1995 Nov 17;270(46):27407–27410. doi: 10.1074/jbc.270.46.27407. [DOI] [PubMed] [Google Scholar]
  32. Xiao S., Rose D. W., Sasaoka T., Maegawa H., Burke T. R., Jr, Roller P. P., Shoelson S. E., Olefsky J. M. Syp (SH-PTP2) is a positive mediator of growth factor-stimulated mitogenic signal transduction. J Biol Chem. 1994 Aug 19;269(33):21244–21248. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES