This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Razzini, G. Articles by Falasca, M. Search for Related Content PubMed PubMed Citation Articles by Razzini, G. Articles by Falasca, M.

Different Subcellular Localization and Phosphoinositides Binding of Insulin Receptor Substrate Protein Pleckstrin Homology Domains

Unit of Physiopathology of Cell Signaling (G.R., A.I., A.B., M.F.) Laboratory of Cellular and Molecular Endocrinology Department of Cell Biology & Oncology Istituto di Ricerche Farmacologiche "Mario Negri" Consorzio Mario Negri Sud, 66030 Santa Maria Imbaro (CH), Italy
Department of Experimental Medicine and Pathology (S.S.) Universita’ "La Sapienza" di Roma 00161 Roma, Italy
Department of Oncology and Neuroscience (D.L.E.) Università G. D’Annunzio Chieti 66100 Chieti, Italy

Insulin evokes diverse biological effects through receptor-mediated tyrosine phosphorylation of the insulin receptor substrate (IRS) proteins. Here, we show that, in vitro, the IRS-1, -2 and -3 pleckstrin homology (PH) domains bind with different specificities to the 3-phosphorylated phosphoinositides. In fact, the IRS-1 PH domain binds preferentially to phosphatidylinositol 3,4,5-trisphosphate (PtdIns-3,4,5-P₃), the IRS-2 PH domain to phosphatidylinositol 3,4-bisphosphate (PtdIns-3,4-P₂), and the IRS-3 PH domain to phosphatidylinositol 3-phosphate. When expressed in NIH-IR fibroblasts and L6 myocytes, the IRS-1 and -2 PH domains tagged with green fluorescent protein (GFP) are localized exclusively in the cytoplasm. Stimulation with insulin causes a translocation of the GFP-IRS-1 and -2 PH domains to the plasma membrane within 3–5 min. This translocation is blocked by the phosphatidylinositol 3-kinase (PI 3-K) inhibitors, wortmannin and LY294002, suggesting that this event is PI 3-K dependent. Interestingly, platelet-derived growth factor (PDGF) did not induce translocation of the IRS-1 and -2 PH domains to the plasma membrane, indicating the existence of specificity for insulin. In contrast, the GFP-IRS-3 PH domain is constitutively localized to the plasma membrane. These results reveal a differential regulation of the IRS PH domains and a novel positive feedback loop in which PI 3-K functions as both an upstream regulator and a downstream effector of IRS-1 and -2 signaling.

This article has been cited by other articles:

				J. Wauman, A.-S. De Smet, D. Catteeuw, D. Belsham, and J. Tavernier Insulin Receptor Substrate 4 Couples the Leptin Receptor to Multiple Signaling Pathways Mol. Endocrinol., April 1, 2008; 22(4): 965 - 977. [Abstract] [Full Text] [PDF]

				R. Nawaratne, A. Gray, C. H. Jorgensen, C. P. Downes, K. Siddle, and J. K. Sethi Regulation of Insulin Receptor Substrate 1 Pleckstrin Homology Domain by Protein Kinase C: Role of Serine 24 Phosphorylation Mol. Endocrinol., August 1, 2006; 20(8): 1838 - 1852. [Abstract] [Full Text] [PDF]

				X. Cui, H.-J. Kim, I. Kuiatse, H. Kim, P. H. Brown, and A. V. Lee Epidermal Growth Factor Induces Insulin Receptor Substrate-2 in Breast Cancer Cells via c-Jun NH2-Terminal Kinase/Activator Protein-1 Signaling to Regulate Cell Migration. Cancer Res., May 15, 2006; 66(10): 5304 - 5313. [Abstract] [Full Text] [PDF]

				K. Chakrabarti, R. Lin, N. I. Schiller, Y. Wang, D. Koubi, Y.-X. Fan, B. B. Rudkin, G. R. Johnson, and M. R. Schiller Critical Role for Kalirin in Nerve Growth Factor Signaling through TrkA Mol. Cell. Biol., June 15, 2005; 25(12): 5106 - 5118. [Abstract] [Full Text] [PDF]

				C. Lassarre and J.-M. Ricort Growth Factor-Specific Regulation of Insulin Receptor Substrate-1 Expression in MCF-7 Breast Carcinoma Cells: Effects on the Insulin-Like Growth Factor Signaling Pathway Endocrinology, November 1, 2003; 144(11): 4811 - 4819. [Abstract] [Full Text] [PDF]

				H. H. Bock, Y. Jossin, P. Liu, E. Forster, P. May, A. M. Goffinet, and J. Herz Phosphatidylinositol 3-Kinase Interacts with the Adaptor Protein Dab1 in Response to Reelin Signaling and Is Required for Normal Cortical Lamination J. Biol. Chem., October 3, 2003; 278(40): 38772 - 38779. [Abstract] [Full Text] [PDF]

				T. Maffucci, G. Razzini, A. Ingrosso, H. Chen, S. Iacobelli, S. Sciacchitano, M. J. Quon, and M. Falasca Role of Pleckstrin Homology Domain in Regulating Membrane Targeting and Metabolic Function of Insulin Receptor Substrate 3 Mol. Endocrinol., August 1, 2003; 17(8): 1568 - 1579. [Abstract] [Full Text] [PDF]

				H. Sun, X. Tu, M. Prisco, A. Wu, I. Casiburi, and R. Baserga Insulin-Like Growth Factor I Receptor Signaling and Nuclear Translocation of Insulin Receptor Substrates 1 and 2 Mol. Endocrinol., March 1, 2003; 17(3): 472 - 486. [Abstract] [Full Text] [PDF]

				X. Tu, P. Batta, N. Innocent, M. Prisco, I. Casaburi, B. Belletti, and R. Baserga Nuclear Translocation of Insulin Receptor Substrate-1 by Oncogenes And Igf-I. EFFECT ON RIBOSOMAL RNA SYNTHESIS* J. Biol. Chem., November 8, 2002; 277(46): 44357 - 44365. [Abstract] [Full Text] [PDF]

				J. Farhang-Fallah, V. K. Randhawa, A. Nimnual, A. Klip, D. Bar-Sagi, and M. Rozakis-Adcock The Pleckstrin Homology (PH) Domain-Interacting Protein Couples the Insulin Receptor Substrate 1 PH Domain to Insulin Signaling Pathways Leading to Mitogenesis and GLUT4 Translocation Mol. Cell. Biol., October 15, 2002; 22(20): 7325 - 7336. [Abstract] [Full Text] [PDF]

				Y.-H. Tseng, K. Ueki, K. M. Kriauciunas, and C. R. Kahn Differential Roles of Insulin Receptor Substrates in the Anti-apoptotic Function of Insulin-like Growth Factor-1 and Insulin J. Biol. Chem., August 23, 2002; 277(35): 31601 - 31611. [Abstract] [Full Text] [PDF]

				S. H. Lee, J. B. Jin, J. Song, M. K. Min, D. S. Park, Y.-W. Kim, and I. Hwang The Intermolecular Interaction between the PH Domain and the C-terminal Domain of Arabidopsis Dynamin-like 6 Determines Lipid Binding Specificity J. Biol. Chem., August 23, 2002; 277(35): 31842 - 31849. [Abstract] [Full Text] [PDF]

				M. Prisco, F. Santini, R. Baffa, M. Liu, R. Drakas, A. Wu, and R. Baserga Nuclear Translocation of Insulin Receptor Substrate-1 by the Simian Virus 40 T Antigen and the Activated Type 1 Insulin-like Growth Factor Receptor J. Biol. Chem., August 23, 2002; 277(35): 32078 - 32085. [Abstract] [Full Text] [PDF]

				T. Kabuta, F. Hakuno, T. Asano, and S.-I. Takahashi Insulin Receptor Substrate-3 Functions as Transcriptional Activator in the Nucleus J. Biol. Chem., February 22, 2002; 277(9): 6846 - 6851. [Abstract] [Full Text] [PDF]

				M. Navarro, B. Valentinis, B. Belletti, G. Romano, K. Reiss, and R. Baserga Regulation of Id2 Gene Expression by the Type 1 IGF Receptor and the Insulin Receptor Substrate-1 Endocrinology, December 1, 2001; 142(12): 5149 - 5157. [Abstract] [Full Text] [PDF]

				L. Simpson, J. Li, D. Liaw, I. Hennessy, J. Oliner, F. Christians, and R. Parsons PTEN Expression Causes Feedback Upregulation of Insulin Receptor Substrate 2 Mol. Cell. Biol., June 15, 2001; 21(12): 3947 - 3958. [Abstract] [Full Text]

				Y. Tsuji, Y. Kaburagi, Y. Terauchi, S. Satoh, N. Kubota, H. Tamemoto, F. B. Kraemer, H. Sekihara, S. Aizawa, Y. Akanuma, et al. Subcellular Localization of Insulin Receptor Substrate Family Proteins Associated With Phosphatidylinositol 3-Kinase Activity and Alterations in Lipolysis in Primary Mouse Adipocytes From IRS-1 Null Mice Diabetes, June 1, 2001; 50(6): 1455 - 1463. [Abstract] [Full Text]

				I. Vainshtein, K. S. Kovacina, and R. A. Roth The Insulin Receptor Substrate (IRS)-1 Pleckstrin Homology Domain Functions in Downstream Signaling J. Biol. Chem., March 9, 2001; 276(11): 8073 - 8078. [Abstract] [Full Text] [PDF]

				A. R. Jacobs, D. LeRoith, and S. I. Taylor Insulin Receptor Substrate-1 Pleckstrin Homology and Phosphotyrosine-binding Domains Are Both Involved in Plasma Membrane Targeting J. Biol. Chem., October 26, 2001; 276(44): 40795 - 40802. [Abstract] [Full Text] [PDF]