The Pleckstrin Homology Domain Is the Principle Link between the Insulin Receptor and IRS-1*

Interaction domains located in the NH2 terminus of IRS-1 mediate its recognition by the insulin receptor. Alignment of IRS-1 and IRS-2 reveals two homology regions: the IH1PH contains a pleckstrin homology (PH) domain, and the IH2PTB contains a phosphotyrosine binding (PTB) domain. A third region in IRS-1 called SAIN was proposed to contain another functional PTB domain. Peptide competition experiments demonstrated that the IH2PTB in IRS-2, like the corresponding domain in IRS-1, binds directly to peptides containing NPXY motifs. In contrast, these peptides do not bind to IH1PH or the SAIN regions. In 32D cells the IH1PH was essential for insulin-stimulated tyrosine phosphorylation of IRS-1 and insulin-stimulated phosphatidylinositol 3-kinase activity and p70s6k phosphorylation. In contrast, the IH2PTB and the SAIN regions were not required for these insulin actions; however, the IH2PTB improved the coupling between IRS-1 and the insulin receptor. Overexpression of the insulin receptor in 32DIR cells increased IRS-1 tyrosine phosphorylation and mediated insulin-stimulated DNA synthesis. The sensitivity of these responses was partially reduced by deletion of either the IH1PH or the IH2PTB and significantly reduced when both regions were deleted together. Thus, the PH and PTB domains equally couple IRS-1 to high levels of insulin receptor normally expressed in most cells, whereas at low levels of insulin receptors the PTB domain is inefficient and the PH domain is essential for a productive interaction.

[1]  G. Crabtree,et al.  Rapamycin-FKBP specifically blocks growth-dependent activation of and signaling by the 70 kd S6 protein kinases , 1992, Cell.

[2]  M. White,et al.  Preparation and use of anti-phosphotyrosine antibodies to study structure and function of insulin receptor. , 1991, Methods in enzymology.

[3]  M. White,et al.  PTB Domains of IRS-1 and Shc Have Distinct but Overlapping Binding Specificities (*) , 1995, The Journal of Biological Chemistry.

[4]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Julie A. Pitcher,et al.  Pleckstrin Homology Domain-mediated Membrane Association and Activation of the -Adrenergic Receptor Kinase Requires Coordinate Interaction with G Subunits and Lipid(*) , 1995, The Journal of Biological Chemistry.

[6]  J. Briscoe,et al.  Signal Transduction: Just another signalling pathway , 1994, Current Biology.

[7]  P. Hajduk,et al.  Pleckstrin homology domains bind to phosphatidylinositol-4,5-bisphosphate , 1994, Nature.

[8]  M. White,et al.  New frontiers in insulin receptor substrate signaling , 1995, Trends in Endocrinology & Metabolism.

[9]  A. Godwin,et al.  A Grb2-associated docking protein in EGF- and insulin-receptor signalling , 1996, Nature.

[10]  T. O’Neill,et al.  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 (*) , 1995, The Journal of Biological Chemistry.

[11]  M. White,et al.  IRS-1: essential for insulin- and IL-4-stimulated mitogenesis in hematopoietic cells. , 1993, Science.

[12]  E. Van Obberghen,et al.  Insulin Receptor Substrate-2 Binds to the Insulin Receptor through Its Phosphotyrosine-binding Domain and through a Newly Identified Domain Comprising Amino Acids 591786 (*) , 1996, The Journal of Biological Chemistry.

[13]  J. Johnston,et al.  Interleukins 2, 4, 7, and 15 Stimulate Tyrosine Phosphorylation of Insulin Receptor Substrates 1 and 2 in T Cells POTENTIAL ROLE OF JAK KINASES (*) , 1995, The Journal of Biological Chemistry.

[14]  M. White,et al.  The Type I Interferon Receptor Mediates Tyrosine Phosphorylation of Insulin Receptor Substrate 2 (*) , 1996, The Journal of Biological Chemistry.

[15]  M. White,et al.  Interferon-α Engages the Insulin Receptor Substrate-1 to Associate with the Phosphatidylinositol 3′-Kinase (*) , 1995, The Journal of Biological Chemistry.

[16]  L. Williams,et al.  An alternative to SH2 domains for binding tyrosine-phosphorylated proteins. , 1994, Science.

[17]  C. Kahn,et al.  Insulin stimulates serine and tyrosine phosphorylation in the juxtamembrane region of the insulin receptor. , 1993, The Journal of biological chemistry.

[18]  C. Kahn,et al.  Structure of the insulin receptor substrate IRS-1 defines a unique signal transduction protein , 1991, Nature.

[19]  S. Shoelson,et al.  Structure of the IRS-1 PTB Domain Bound to the Juxtamembrane Region of the Insulin Receptor , 1996, Cell.

[20]  Peer Bork,et al.  A phosphotyrosine interaction domain , 1995, Cell.

[21]  M. Nakafuku,et al.  Signal transduction pathways from insulin receptors to Ras. Analysis by mutant insulin receptors. , 1994, The Journal of biological chemistry.

[22]  William Arbuthnot Sir Lane,et al.  Role of IRS-2 in insulin and cytokine signalling , 1995, Nature.

[23]  J. Blenis,et al.  The Pleckstrin Homology Domain in Insulin Receptor Substrate-1 Sensitizes Insulin Signaling (*) , 1995, The Journal of Biological Chemistry.

[24]  R. Lefkowitz,et al.  Gβγ interactions with PH domains and Ras-MAPK signaling pathways , 1995 .

[25]  M. White,et al.  Insulin receptor substrate 1 rescues insulin action in CHO cells expressing mutant insulin receptors that lack a juxtamembrane NPXY motif , 1995, Molecular and cellular biology.

[26]  A. Craparo,et al.  Phosphotyrosine-dependent interaction of SHC and insulin receptor substrate 1 with the NPEY motif of the insulin receptor via a novel non-SH2 domain , 1995, Molecular and cellular biology.

[27]  M. White,et al.  The insulin receptor substrate-1-related 4PS substrate but not the interleukin-2R gamma chain is involved in interleukin-13-mediated signal transduction. , 1995, Blood.

[28]  C. Turck,et al.  PTB domain binding to signaling proteins through a sequence motif containing phosphotyrosine. , 1995, Science.

[29]  A. Ullrich,et al.  Mutation of the insulin receptor at tyrosine 960 inhibits signal transmission but does not affect its tyrosine kinase activity , 1988, Cell.

[30]  Tony Pawson,et al.  Protein modules and signalling networks , 1995, Nature.

[31]  G. Lienhard,et al.  Common elements in interleukin 4 and insulin signaling pathways in factor-dependent hematopoietic cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[32]  J. Schlessinger,et al.  PH Domains: Diverse Sequences with a Common Fold Recruit Signaling Molecules to the Cell Surface , 1996, Cell.

[33]  Role of IRS-1-GRB-2 complexes in insulin signaling. , 1994, Molecular and cellular biology.

[34]  C. Kahn,et al.  The insulin signaling system. , 1994, The Journal of biological chemistry.

[35]  B. Neel,et al.  SH-PTP2/Syp SH2 domain binding specificity is defined by direct interactions with platelet-derived growth factor beta-receptor, epidermal growth factor receptor, and insulin receptor substrate-1-derived phosphopeptides. , 1994, The Journal of biological chemistry.

[36]  A. Petros,et al.  Structure and ligand recognition of the phosphotyrosine binding domain of Shc , 1995, Nature.

[37]  J. Blenis,et al.  The Drosophila insulin receptor activates multiple signaling pathways but requires insulin receptor substrate proteins for DNA synthesis , 1996, Molecular and cellular biology.

[38]  M. White,et al.  Growth Hormone, Interferon-γ, and Leukemia Inhibitory Factor Promoted Tyrosyl Phosphorylation of Insulin Receptor Substrate-1 (*) , 1995, The Journal of Biological Chemistry.

[39]  M J Saad,et al.  Regulation of insulin receptor substrate-1 in liver and muscle of animal models of insulin resistance. , 1992, The Journal of clinical investigation.