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Rapid Publication

The Gene INPPL1, Encoding the Lipid Phosphatase SHIP2, Is a Candidate for Type 2 Diabetes In Rat and Man

  1. Evelyne Marion1,
  2. Pamela Jane Kaisaki2,
  3. Valérie Pouillon1,
  4. Cyril Gueydan3,
  5. Jonathan C. Levy4,
  6. André Bodson5,
  7. Georges Krzentowski5,
  8. Jean-Claude Daubresse6,
  9. Jean Mockel7,
  10. Jens Behrends8,
  11. Geneviève Servais9,
  12. Claude Szpirer10,
  13. Véronique Kruys3,
  14. Dominique Gauguier2 and
  15. Stéphane Schurmans1
  1. 1IRIBHM (Institut de Recherches en Biologie Humaine et Moléculaire), IBMM (Institut de Biologie et de Médecine Moléculaires), ULB (Université Libre de Bruxelles), Gosselies, Belgium
  2. 2Wellcome Trust Centre for Human Genetics, University of Oxford, Headington, U.K.
  3. 3Laboratoire de Chimie Biologique, IBMM, ULB, Gosselies, Belgium
  4. 4Diabetes Research Laboratories, University of Oxford, Headington, U.K.
  5. 5Department of Internal Medicine, Endocrinology-Diabetology, C.H.U. (Centre Hospitalier Universitaire) de Charleroi, site de Jumet, Jumet, Belgium
  6. 6Department of Internal Medicine, Endocrinology-Diabetology, C.H.U. de Charleroi, Charleroi, Belgium
  7. 7Department of Endocrinology, Hopital Erasme, ULB, Brussels, Belgium
  8. 8Department of Clinical Endocrinology, Medical School Hannover, Germany
  9. 9Department of Immunology, C.H.U. Brugmann-Huderf, ULB, Brussels, Belgium
  10. 10Laboratoire de Biologie du Développement, IBMM, ULB, Gosselies, Belgium
    Diabetes 2002 Jul; 51(7): 2012-2017. https://doi.org/10.2337/diabetes.51.7.2012
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    • FIG. 1
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      FIG. 1

      The R1142C mutation in GK and SH rat strains: detection and analysis. A: Localization of the SHIP2 gene and the Nidd/gk1(GK rat) QTL on the long arm of rat chromosome 1. B: The R1142C mutation found in the GK and SH rat strains. C: Akt/PKB and MAP kinase activities in CHO-IR stimulated or not for 2 min with 10 nmol/l insulin. Results are the means of triplicate ± SE and are representative of a typical experiment out of three. Western blot analysis shows that the same amount of wild-type and mutated HA-tagged SHIP2 was expressed after transfection. *P < 0.05; **P < 0.02; ***P < 0.002; ****P < 10-4 (by Student�s t test).

    • FIG. 2
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      FIG. 2

      The 3'UTR of the SHIP2 gene: structure and mutations. A: The Δ16-bp mutation found in the SHIP2 3'UTR from the SMH10 subject: sequence of the PCR product amplified from SMH10 cDNA (above), and sequences of the wild-type (middle) and the Δ16-bp mutant (below) after subcloning. B and C: Structure of the SHIP2 3'UTR: number and location of ATTTA pentamers and ATTTTA hexamers in man, rat, and mouse. Definition of the three mutations (Δ4bp, G/A substitution, and Δ16 bp) found in the SHIP2 3'UTR, and location of the regions with a high percentage of identity between mouse, rat, and man.

    • Fig. 3
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      Fig. 3

      Effects of the wild-type and the Δ16 SHIP2 3' UTRs on luciferase reporter gene expression. Transfection efficiencies were assessed by β-galactidose protein quantification, and luciferase reporter gene expression was corrected accordingly. Differences in transfection efficiencies between the tested plasmids of the same experiment were always <10%. A: Luciferase expression in rat L6 cells transfected with a plasmid vector containing the 3'UTR from SV40, rat, mouse, or human SHIP2 gene. Results are the means ± SE and are representative of three separate experiments performed in triplicate. *P < 0.03; **P < 0.005 (Student's t test). B: Luciferase expression in human 293 cells transfected with a plasmid vector containing either the wild-type or the Δ16-bp mutation. Results are the means ± SE and are representative of four separate experiments performed in triplicate. C: Total RNA isolated from human 293 cells transfected with the empty plasmid vector (0) or a plasmid vector containing either the wild-type (WT) or the Δ16-bp mutated (Δ16) SHIP2 was analyzed with a luciferase or an actin probe. Densitometric analysis of the signals was performed with a phosphorimager. Results are the means ± SE.

    Tables

    • Figures
    • TABLE 1

      cDNA sequence analysis around amino acid 1142 (SHIP2) or the corresponding SHIP1 cDNA region in various species

      NameAccession number1142
      Class II ligand consensusPXXPXR
      Rat SHIP2AB011439P-GPGRSALLP
      Rat SHIP2AB025794P-GPGRSALLP
      Brown-Norway rat SHIP2*�P-GPGRSALLP
      Zucker-Fatty rat SHIP2�P-GPGRSALLP
      Goto-Kakizaki rat SHIP2�P-GPGCSALLP
      SHR rat SHIP2§�P-GPGCSALLP
      Mouse SHIP2AF162781P-GPGRSALLP
      Human SHIP2Y14385PAGPARSALLP
      Rat SHIP1U55192PVKPSRSEMSQ
      Mouse SHIP1NM010566PVKPSRSEMSQ
      Human SHIP1U57650PIKPSRSEINQ
      • *

        * As F344, COP, WF, WKY, WKY Leicester, SD, PD, Buffalo, Mib, LEW, BKI, BDII, LE, MNS, SS/jr, SR/jr and SPRD rat strains;

      • §

        § includes SHR/le, SHRSP/gla, and the original strain SHRSP/izm.

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    The Gene INPPL1, Encoding the Lipid Phosphatase SHIP2, Is a Candidate for Type 2 Diabetes In Rat and Man
    Evelyne Marion, Pamela Jane Kaisaki, Valérie Pouillon, Cyril Gueydan, Jonathan C. Levy, André Bodson, Georges Krzentowski, Jean-Claude Daubresse, Jean Mockel, Jens Behrends, Geneviève Servais, Claude Szpirer, Véronique Kruys, Dominique Gauguier, Stéphane Schurmans
    Diabetes Jul 2002, 51 (7) 2012-2017; DOI: 10.2337/diabetes.51.7.2012

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    The Gene INPPL1, Encoding the Lipid Phosphatase SHIP2, Is a Candidate for Type 2 Diabetes In Rat and Man
    Evelyne Marion, Pamela Jane Kaisaki, Valérie Pouillon, Cyril Gueydan, Jonathan C. Levy, André Bodson, Georges Krzentowski, Jean-Claude Daubresse, Jean Mockel, Jens Behrends, Geneviève Servais, Claude Szpirer, Véronique Kruys, Dominique Gauguier, Stéphane Schurmans
    Diabetes Jul 2002, 51 (7) 2012-2017; DOI: 10.2337/diabetes.51.7.2012
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