Genetic ablation of Inppl1, which encodes SHIP2 (SH2-domain containing inositol 5-phosphatase 2), was previously reported to induce severe insulin sensitivity, leading to early postnatal death. In the previous study, the targeting construct left the first eighteen exons encoding Inppl1 intact, generating a Inppl1EX19-28−/− mouse, and apparently also deleted a second gene, Phox2a. We report a new SHIP2 knockout (Inppl1−/−) targeted to the translation-initiating ATG, which is null for Inppl1 mRNA and protein. Inppl1−/− mice are viable, have normal glucose and insulin levels, and normal insulin and glucose tolerances. The Inppl1−/− mice are, however, highly resistant to weight gain when placed on a high-fat diet. These results suggest that inhibition of SHIP2 would be useful in the effort to ameliorate diet-induced obesity, but call into question a dominant role of SHIP2 in modulating glucose homeostasis.
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House, P.D. & Weidemann, M.J. Characterization of an [125I]-insulin binding plasma membrane fraction from rat liver. Biochem. Biophys. Res. Commun. 41, 541–548 (1970).
Freychet, P., Roth, J. & Neville, D.M.J. Insulin receptors in the liver: specific binding of (125I)insulin to the plasma membrane and its relation to insulin bioactivity. Proc. Natl. Acad. Sci. USA 68, 1833–1837 (1971).
Cuatrecasas, P. Properties of the insulin receptor of isolated fat cell membranes. J. Biol. Chem. 246, 7265–7274 (1971).
Kasuga, M., Karlsson, F.A. & Kahn, C.R. Insulin stimulates the phosphorylation of the 95,000-Dalton subunit of its own receptor. Science 215, 185–187 (1982).
Asante-Appiah, E. & Kennedy, B.P. Protein tyrosine phosphatases: the quest for negative regulators of insulin action. Am. J. Physiol. Endocrinol. Metab. 284, E663–E670 (2003).
Elchebly, M. et al. Increased insulin sensitivity and obesity resistance in mice lacking the protein tyrosine phosphatase-1b gene. Science 283, 1544–1548 (1999).
Klaman, L.D. et al. Increased energy expenditure, decreased adiposity, and tissue-specific insulin sensitivity in protein-tyrosine phosphatase 1b-deficient mice. Mol. Cell. Biol. 20, 5479–5489 (2000).
Sun, X.J. et al. Structure of the insulin receptor substrate IRS-1 defines a unique signal transduction protein. Nature 352, 73–77 (1991).
Hadari, Y.R. et al. Insulin and insulinomimetic agents induce activation of phosphatidylinositol 3-kinase upon its association with pp185 (IRS-1) in intact rat livers. J. Biol. Chem. 267, 17483–17486 (1992).
Burgering, B.M. & Coffer, P.J. Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction. Nature 376, 599–602 (1995).
Franke, T.F. et al. The protein kinase encoded by the Akt proto-oncogene is a target of the PDGF-activated phosphatidylinositol 3-kinase. Cell 81, 727–736 (1995).
Whitman, M., Downes, C.P., Keeler, M., Keller, T. & Cantley, L. Type I phosphatidylinositol kinase makes a novel inositol phospholipid, phosphatidylinositol-3-phosphate. Nature 332, 644–646 (1988).
Toker, A. & Cantley, L.C. Signalling through the lipid products of phosphoinositide-3-OH kinase. Nature 387, 673–676 (1997).
Alessi, D.R. et al. Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activates protein kinase B alpha. Curr. Biol. 7, 261–269 (1997).
Andjelkovic, M. et al. Role of translocation in the activation and function of protein kinase B. J. Biol. Chem. 272, 31515–31524 (1997).
Andjelkovic, M. et al. Activation and phosphorylation of a pleckstrin homology domain containing protein kinase (RAC-PK/PKB) promoted by serum and protein phosphatase inhibitors. Proc. Natl. Acad. Sci. USA 93, 5699–5704 (1996).
Chen, W.S. et al. Growth retardation and increased apoptosis in mice with homozygous disruption of the Akt1 gene. Genes Dev. 15, 2203–2208 (2001).
Cho, H. et al. Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta). Science 292, 1728–1731 (2001).
Katome, T. et al. Use of RNA-interference-mediated gene silencing and adenoviral overexpression to elucidate the roles of AKT/PKB-isoforms in insulin actions. J. Biol. Chem. 278, 28312–28323 (2003).
Summers, S.A., Garza, L.A., Zhou, H. & Birnbaum, M.J. Regulation of insulin-stimulated glucose transporter GLUT4 translocation and Akt kinase activity by ceramide. Mol. Cell. Biol. 18, 5457–5464 (1998).
Hill, M.M. et al. A role for protein kinase Bbeta/Akt2 in insulin-stimulated GLUT4 translocation in adipocytes. Mol. Cell. Biol. 19, 7771–7781 (1999).
Sakoda, G. et al. Differing roles of Akt and serum- and glucocorticoid-regulated kinase in glucose metabolism, DNA synthesis, and oncogenic activity. J. Biol. Chem. 278, 25802–25807 (2003).
Brunet, A. et al. Protein kinase SGK mediates survival signals by phosphorylating the forkhead transcription factor FKHRL1 (FOXO3a). Mol. Cell. Biol. 21, 952–965 (2001).
Hertweck, M., Gobel, C. & Baumeister, R. C. elegans SGK-1 is the critical component in the Akt/PKB kinase complex to control stress response and life span. Dev. Cell 6, 577–588 (2004).
Damen, J.E. et al. The 145-kDa protein induced to associate with Shc by multiple cytokines is an inositol tetraphosphate and phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase. Proc. Natl. Acad. Sci. USA 93, 1689–1693 (1996).
Stambolic, V. et al. Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell 95, 29–39 (1998).
Liu, L. et al. The Src homology 2 (SH2) domain of SH2-containing inositol phosphatase (SHIP) is essential for tyrosine phosphorylation of SHIP, its association with Shc, and its induction of apoptosis. J. Biol. Chem. 272, 8983–8988 (1997).
Lamkin, T.D. et al. Shc interaction with Src homology 2 domain containing inositol phosphatase (SHIP) in vivo requires the Shc-phosphotyrosine binding domain and two specific phosphotyrosines on SHIP. J. Biol. Chem. 272, 10396–401 (1997).
Pesesse, X., Deleu, S., De Smedt, F., Drayer, L. & Erneux, C. Identification of a second SH2-domain-containing protein closely related to the phosphatidylinositol polyphosphate 5-phosphatase SHIP. Biochem. Biophys. Res. Commun. 239, 697–700 (1997).
Pesesse, X. et al. The SH2 domain containing inositol 5-phosphatase SHIP2 displays phosphatidylinositol 3,4,5-trisphosphate and inositol 1,3,4,5- tetrakisphosphate 5-phosphatase activity. FEBS Lett. 437, 301–303 (1998).
Aman, M.J., Lamkin, T.D., Okada, H., Kurosaki, T. & Ravichandran, K.S. The inositol phosphatase SHIP inhibits Akt/PKB activation in B cells. J. Biol. Chem. 273, 33922–33928 (1998).
Clement, S. et al. The lipid phosphatase SHIP2 controls insulin sensitivity. Nature 409, 92–97 (2001).
Wada, T. et al. Overexpression of SH2-Containing Inositol Phosphatase 2 Results in Negative Regulation of Insulin-Induced Metabolic Actions in 3T3-L1 Adipocytes via Its 5-Phosphatase Catalytic Activity. Mol. Cell. Biol. 21, 1633–1646 (2001).
Alessi, D.R. et al. 3-Phosphoinositide-dependent protein kinase-1 (PDK1): structural and functional homology with the Drosophila DSTPK61 kinase. Curr. Biol. 7, 776–789 (1997).
Rommel, C. et al. Mediation of IGF-1-induced skeletal myotube hypertrophy by PI(3)K/Akt/mTOR and PI(3)K/Akt/GSK3 pathways. Nat. Cell Biol. 3, 1009–1013 (2001).
Bodine, S.C. et al. Akt/mTOR pathway is a crucial regulator of skeletal muscle hypertrophy and can prevent muscle atrophy in vivo. Nat. Cell Biol. 3, 1014–1019 (2001).
Boss, O. et al. Uncoupling protein-3: a new member of the mitochondrial carrier family with tissue-specific expression. FEBS Lett. 408, 39–42 (1997).
Clement, S. et al. Corrigendum: The lipid phosphatase SHIP2 controls insulin sensitivity. Nature 431, 878 (2004).
Valenzuela, D. et al. High-throughput engineering of the mouse genome coupled with high-resolution expression analysis. Nat. Biotechnol. 21, 652–659 (2003).
Sleeman, M.W. et al. Ciliary neurotrophic factor improves diabetic parameters and hepatic steatosis and increases basal metabolic rate in db/db mice. Proc. Natl. Acad. Sci. USA 100, 14297–14302 (2003).
We thank L.S. Schleifer and P.R. Vagelos for support, along with the rest of the Regeneron community, and are greatly indebted to our colleagues at Procter & Gamble Pharmaceuticals for their continued support; we also thank B. Ephraim and V. Lan for graphics work, A. Steuernagel for discussions and L. Suva for X-ray analysis. Thank you also to T. Dechiara, W. Poueymirou and M. Simmons for the coordinated breeding of knockout mice.
The authors are stockholders in Regeneron Pharmaceuticals, which could receive financial gains as a result of this publication.
Gene targeting of the SHIP2 locus. (PDF 60 kb)
Facial and X-ray analysis of SHIP2−/− mice did not reveal any gross abnormality. (PDF 96 kb)
SHIP2 deletion protects female mice from diet-induced obesity and insulin resistance. (PDF 111 kb)
Additional analysis of insulin signaling of SHIP2−/− mice, both on regular chow and on a high fat diet. (PDF 352 kb)
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