Summary
The purpose of these experiments was to test the hypothesis that impaired glucose-stimulated insulin secretion in NIDDM is due to mutations in the islet beta cell/liver glucose transporter (GLUT 2) gene. Using oligonucleotide primers flanking each of the 11 exons, the structural portion of the gene was studied by PCR-SSCP analysis. DNA from African-American females (n=48), who had gestational diabetes but developed overt NIDDM after delivery, was studied. Each SSCP variant was sequenced directly from genomic DNA. Two amino acid substitutions from the previously reported sequence were found, one in exon 3 and the other in exon 4 B. Four additional silent mutations in the coding region, and six intron mutations outside the splice junction consensus sequences, were also identified. The mutation GTC x ATC in exon 4B substituted Val197 to Ile197. This amino acid substitution was found in only one NIDDM patient in a single allele, and was not found in 52 control subjects. This residue exists in the fifth membrane spanning domain, and Val at this position is conserved in mouse and rat GLUT 2, and human GLUT 1 to GLUT 4. The other codon change in exon 3, ACT x ATT, substituted Thr110 to Ile110 in the second membrane spanning domain. To determine the frequency of this non-conservative amino acid substitution, a PCR-LCR assay was developed. This assay was simple and highly specific for detection of this single nucleotide substitution. The allelic frequency of the ATT (Ile110) in NIDDM patients (39.6%, n=48) and that in controls (47.1%, n=52) did not differ (p=0.32, Fisher's exact test). In conclusion, we identified two variant GLUT 2 glucose transporters in a subset of NIDDM patients. The rare variant in exon 4 B may contribute to the diabetic susceptibility and awaits further investigation. However, structural abnormalities of the GLUT 2 transporter associated with NIDDM appeared to be rare and were not likely to be a major determinant of genetic susceptibility to this type of diabetes in the population studied.
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Abbreviations
- NIDDM:
-
Non-insulin-dependent diabetes mellitus
- GLUT 2:
-
islet beta cell/liver glucose transporter
- PCR:
-
polymerase chain reaction
- SSCP:
-
single strand conformation polymorphism
- LCR:
-
ligase chain reaction
- bp:
-
base pair
References
DeFronzo RA (1988) Triumvirate: Beta-cell, muscle, liver: A collusion responsible for NIDDM. Diabetes 37:667–687
Barnett AT, Eff C, Leslie RDG, Pyke DA (1981) Diabetes in identical twins. A study of 200 pairs. Diabetologia 20:87–93
Knowler WC, Pettitt DJ, Saad MF, Bennett PH (1990) Diabetes mellitus in the Pima Indians: incidence, risk factors and pathogenesis. Diabetes Metab Rev 6:1–27
Kawate R, Yamakido M, Nishimoto Y, Bennett PH, Hamman RF, Knowler WC (1979) Diabetes mellitus and its vascular complications in Japanese migrants on the island of Hawaii. Diabetes Care 2:161–170
Steiner DF, Tager HS, Chan SJ, Nanjo K, Sanke T, Rubenstein AH (1990) Lessons learned from molecular biology of insulin-gene mutations. Diabetes Care 13:660–669
Olansky L, Welling C, Giddings S et al. (1992) A variant insulin promoter in non-insulin-dependent diabetes mellitus. J Clin Invest 89:1596–1602
Taylor S, Cama A, Accili D et al. (1991) Genetic basis of endocrine disease 1. Molecular genetics of insulin resistant diabetes mellitus. J Clin Endocrinol Metab 73:1158–1163
Ballinger SW, Shoffner JM, Hedaya EV et al. (1992) Maternally transmitted diabetes and deafness associated with a 10.4 kb mitochondrial deletion. Nature Genetics 1: 11–15
Ouweland JMW, Lemkes HHPJ, Ruitenbeek W et al. (1992) Mutation in mitochondrial tRNALeu(UUR) gene in a large pedigree with maternally transmitted type II diabetes mellitus and deafness. Nature Genetics 1:368–371
Reardon W, Ross RJM, Sweeney MG et al. (1992) Diabetes mellitus associated with a pathogenic point mutation in mitochondrial DNA. Lancet 340:1376–1379
Froguel Ph, Vaxillaire M, Sun F et al. (1992) Close linkage of glucokinase locus on chromosome 7 p to early-onset non-insulin-dependent diabetes mellitus. Nature 356:162–164
Hattersley AT, Turner RC, Permutt MA et al. (1992) Linkage of type 2 diabetes to the glucokinase gene. Lancet 339: 1307–1310
Froguel Ph, Zouali H, Vionnet N et al. (1993) Familial hyperglycemia due to mutations in glucokinase: definition of a subtype of diabetes mellitus. N Engl J Med 328:697–702
Permutt MA, Chiu KC, Tanizawa Y (1992) Glucokinase and NIDDM: a candidate gene that paid off. Diabetes 41:1367–1372
Thorens B, Sarkar HK, Kaback HR, Lodish HF (1988) Cloning and functional expression in bacteria of a novel glucose transporter present in liver, intestine, kidney and β-pancreatic islet cells. Cell 55:281–290
Permutt MA, Koranyi L, Keller K, Lacy PE, Sharp DW, Mueckler M (1989) Cloning and functional expression of a human pancreatic islet glucose transporter cDNA. Proc Natl Acad Sci USA 86:8688–8692
Thorens B (1992) Molecular and cellular physiology of GLUT-2, a high-Km facilitated diffusion glucose transporter. Internatl Rev Cytol 137 A: 209–238
Newgard CB, Quaade C, Hughes SD, Milburn JL (1990) Glucokinase and glucose transporter expression in liver and islets: implications for control of glucose homeostasis. Biochem Soc Trans 18:851–853
Unger RH (1991) Diabetic hyperglycemia: link to impaired glucose transport in pancreatic β cell. Science 251:1200–1205
Matschinsky FM (1990) Glucokinase as glucose sensor and metabolic signal generator in pancreatic beta-cell and hepatocytes. Diabetes 39:647–652
Tal M, Liang Y, Najafji H, Lodish H, Matschinsky FM (1992) Expression and function of GLUT-1 and GLUT-2 glucose transporter isoforms in cells of cultured rat pancreatic islets. J Biol Chem 267:17241–17247
Matsutani A, Koranyi L, Cox N, Permutt MA (1990) Polymorphisms of GLUT2 and GLUT4 genes. Use in evaluation of genetic susceptibility to NIDDM in Blacks. Diabetes 39: 1534–1542
Patel P, Bell GI, Cook JTE, Turner RC, Wainscoat JS (1991) Multiple restriction fragment length polymorphisms at the GLUT 2 locus: GLUT 2 haplotypes for genetic analysis of type 2 (non-insulin-dependent) diabetes mellitus. Diabetologia 34:817–821
Baroni MG, Alcolado JC, Pozzilli P, Cavallo MG, Li S-R, Galton DJ (1992) Polymorphisms at the GLUT2 (β- cell/liver) glucose transporter gene and non-insulin-dependent diabetes mellitus (NIDDM): analysis in affected pedigree members. Clin Genet 41:229–234
Patel P, Lo YMD, Hattersley A et al. (1992) Linkage analysis of maturity-onset diabetes of the young with microsatellite polymorphisms. No linkage to ADA to or GLUT2 genes in two families. Diabetes 41:962–967
Elbein SC, Hoffman MD, Matsutani A, Permutt MA (1992) Linkage analysis of GLUT1 (HepG2) and GLUT2 (liver/islet) genes in familial NIDDM. Diabetes 41:1660–1667
Alcolado JC, Baroni MG, Li SR (1991) Association between a restriction fragment length polymorphism at the liver/islet cell (GLUT2) glucose transporter and familial type 2 (non-insulin-dependent) diabetes mellitus. Diabetologia 34:734–736
Cox NJ, Bell GI (1989) Disease associations: chance, artifact, or susceptibility genes? Diabetes 38:947–950
O'Rahilly S, Wainscoat JS, Turner RC (1988) Type 2 (non-insulin-dependent) diabetes mellitus: new genetics for old nightmares. Diabetologia 31:407–414
Lander ES, Botstein D (1986) Strategies for studying heterogeneous genetic traits in human by using a linkage map of restriction fragment length polymorphisms. Proc Natl Acad Sci USA 83:7353–7357
Orita M, Suzuki Y, Sekiya T, Hayashi K (1989) Rapid and sensitive detection of point mutation and DNA polymorphisms using the polymerase chain reaction. Genomics 5: 874–879
Chiu KC, Go RCP, Aoki M, Riggs AC, Tanizawa Y, Acton RT et al. (1994) Glucokinase gene in gestational diabetes mellitus: population association study and molecular scanning. Diabetologia 37:104–110
Acton RT, Vanichanan CJ, Perkins L et al. (1990) Immunogenetic predictors of gestational diabetes in American Blacks. In: Andreani D, Bompiani GD, Di Mario U, Faulk WP, Galluzzo A (eds), Immunobiology of normal and diabetic pregnancy. John Wiley & Sons, Ltd., New York, pp 207–220
Kunkel LM, Smith RD, Soyer SH et al. (1977) Analysis of human Y chromosome specific reiterated DNA in chromosome variants. Proc Natl Acad Sci USA 74: 1245–1249
Chiu KC, Tanizawa Y, Permutt MA (1993) Glucokinase gene variants in the common form of NIDDM. Diabetes 42: 579–582
Matsutani A, Hing A, Steinbrueck T et al. (1992) Mapping the human liver/islet glucose transporter (GLUT2) gene within a genetic linkage map of chromosome 3 q using a (CA)n dinucleotide repeat polymorphism and characterization of the polymorphism in three racial groups. Genomics 13:495–501
Barany F (1991) Genetic disease detection and DNA amplification using cloned thermostable ligase. Proc Natl Acad Sci USA 88:189–193
Fukumoto H, Seino S, Imura H et al. (1988) Sequence, tissue distribution, and chromosomal localization of mRNA encoding a human glucose transporter-like protein. Proc Natl Acad Sci USA 85:5434–5438
Takeda J, Kayano T, Fukumoto H, Bell GI (1993) Organization of the human GLUT2 (pancreatic β-cell and hepatocyte) glucose transporter gene. Diabetes 42: 773–777
Cousins L (1991) Insulin sensitivity in pregnancy. Diabetes 40 [Suppl 2]: 39–43
Kühl C (1991) Insulin secretion and insulin resistance in pregnancy and GDM. Implications for diagnosis and management. Diabetes 40 [Suppl 2]: 18–24
Bell GI, Kayano T, Buse JB et al. (1990) Molecular biology of mammalian glucose transporters. Diabetes Care 13:198–208
Keller K, Strube M, Mueckler M (1990) Functional expression of the human HepG2 and rat adipocyte glucose transporters in Xenopus oocytes. Comparison of kinetic parameters. J Biol Chem 264: 18884–18889
Janssen R, Bogardus C, Takeda J, Thompson DB (1993) A threonine for isoleucine substitution in a membrane spanning domain of the human islet/liver glucose transporter gene. Diabetes 42 [Suppl 1]: 110 A (Abstract)
Jeunemaitre X, Soubrier F, Koteletsev YV et al. (1992) Molecular basis of human hypertension: role of angiotensigen. Cell 71:169–180
Elbein SC, Sorensen LK, Schumacher MC (1993) Methionine for valine substitution in exon 17 of the insulin receptor gene in a pedigree with familial NIDDM. Diabetes 42: 429–434
Cotton RGH (1989) Detection of single base change in nucleic acid. Biochem J 263:1–10
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Tanizawa, Y., Riggs, A.C., Chiu, K.C. et al. Variability of the pancreatic islet beta cell/liver (GLUT 2) glucose transporter gene in NIDDM patients. Diabetologia 37, 420–427 (1994). https://doi.org/10.1007/BF00408481
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DOI: https://doi.org/10.1007/BF00408481