The Gene INPPL1, Encoding the Lipid Phosphatase SHIP2, Is a Candidate for Type 2 Diabetes In Rat and Man
Article Figures & Tables
Figures
- 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
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
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
- TABLE 1
cDNA sequence analysis around amino acid 1142 (SHIP2) or the corresponding SHIP1 cDNA region in various species
Name Accession number 1142 Class II ligand consensus PXXPX R Rat SHIP2 AB011439 P-GPG R SALLP Rat SHIP2 AB025794 P-GPG R SALLP Brown-Norway rat SHIP2* � P-GPG R SALLP Zucker-Fatty rat SHIP2 � P-GPG R SALLP Goto-Kakizaki rat SHIP2 � P-GPG C SALLP SHR rat SHIP2§ � P-GPG C SALLP Mouse SHIP2 AF162781 P-GPG R SALLP Human SHIP2 Y14385 PAGPA R SALLP Rat SHIP1 U55192 PVKPS R SEMSQ Mouse SHIP1 NM010566 PVKPS R SEMSQ Human SHIP1 U57650 PIKPS R SEINQ - *
* As F344, COP, WF, WKY, WKY Leicester, SD, PD, Buffalo, Mib, LEW, BKI, BDII, LE, MNS, SS/jr, SR/jr and SPRD rat strains;
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§ includes SHR/le, SHRSP/gla, and the original strain SHRSP/izm.
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