Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 Jul 1;102(1):232–241. doi: 10.1172/JCI2242

Insulin promoter factor-1 gene mutation linked to early-onset type 2 diabetes mellitus directs expression of a dominant negative isoprotein.

D A Stoffers 1, V Stanojevic 1, J F Habener 1
PMCID: PMC509085  PMID: 9649577

Abstract

The homeodomain transcription factor insulin promoter factor-1 (IPF-1) is required for development of the pancreas and also mediates glucose-responsive stimulation of insulin gene transcription. Earlier we described a human subject with pancreatic agenesis attributable to homozygosity for a cytosine deletion in codon 63 of the IPF-1 gene (Pro63fsdelC). Pro63fsdelC resulted in the premature truncation of an IPF-1 protein which lacked the homeodomain required for DNA binding and nuclear localization. Subsequently, we linked the heterozygous state of this mutation with type 2 diabetes mellitus in the extended family of the pancreatic agenesis proband. In the course of expressing the mutant IPF-1 protein in eukaryotic cells, we detected a second IPF-1 isoform, recognized by COOH- but not NH2-terminal-specific antisera. This isoform localizes to the nucleus and retains DNA-binding functions. We provide evidence that internal translation initiating at an out-of-frame AUG accounts for the appearance of this protein. The reading frame crosses over to the wild-type IPF-1 reading frame at the site of the point deletion just carboxy proximal to the transactivation domain. Thus, the single mutated allele results in the translation of two IPF-1 isoproteins, one of which consists of the NH2-terminal transactivation domain and is sequestered in the cytoplasm and the second of which contains the COOH-terminal DNA-binding domain, but lacks the transactivation domain. Further, the COOH-terminal mutant IPF-1 isoform does not activate transcription and inhibits the transactivation functions of wild-type IPF-1. This circumstance suggests that the mechanism of diabetes in these individuals may be due not only to reduced gene dosage, but also to a dominant negative inhibition of transcription of the insulin gene and other beta cell-specific genes regulated by the mutant IPF-1.

Full Text

The Full Text of this article is available as a PDF (571.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baniahmad A., Tsai S. Y., O'Malley B. W., Tsai M. J. Kindred S thyroid hormone receptor is an active and constitutive silencer and a repressor for thyroid hormone and retinoic acid responses. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10633–10637. doi: 10.1073/pnas.89.22.10633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Carty M. D., Lillquist J. S., Peshavaria M., Stein R., Soeller W. C. Identification of cis- and trans-active factors regulating human islet amyloid polypeptide gene expression in pancreatic beta-cells. J Biol Chem. 1997 May 2;272(18):11986–11993. doi: 10.1074/jbc.272.18.11986. [DOI] [PubMed] [Google Scholar]
  3. Finegood D. T., Scaglia L., Bonner-Weir S. Dynamics of beta-cell mass in the growing rat pancreas. Estimation with a simple mathematical model. Diabetes. 1995 Mar;44(3):249–256. doi: 10.2337/diab.44.3.249. [DOI] [PubMed] [Google Scholar]
  4. German M. S., Moss L. G., Wang J., Rutter W. J. The insulin and islet amyloid polypeptide genes contain similar cell-specific promoter elements that bind identical beta-cell nuclear complexes. Mol Cell Biol. 1992 Apr;12(4):1777–1788. doi: 10.1128/mcb.12.4.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gunnery S., Mäivali U., Mathews M. B. Translation of an uncapped mRNA involves scanning. J Biol Chem. 1997 Aug 22;272(34):21642–21646. doi: 10.1074/jbc.272.34.21642. [DOI] [PubMed] [Google Scholar]
  6. Habener J. F., Stoffers D. A. A newly discovered role of transcription factors involved in pancreas development and the pathogenesis of diabetes mellitus. Proc Assoc Am Physicians. 1998 Jan-Feb;110(1):12–21. [PubMed] [Google Scholar]
  7. Horikawa Y., Iwasaki N., Hara M., Furuta H., Hinokio Y., Cockburn B. N., Lindner T., Yamagata K., Ogata M., Tomonaga O. Mutation in hepatocyte nuclear factor-1 beta gene (TCF2) associated with MODY. Nat Genet. 1997 Dec;17(4):384–385. doi: 10.1038/ng1297-384. [DOI] [PubMed] [Google Scholar]
  8. Jonsson J., Carlsson L., Edlund T., Edlund H. Insulin-promoter-factor 1 is required for pancreas development in mice. Nature. 1994 Oct 13;371(6498):606–609. doi: 10.1038/371606a0. [DOI] [PubMed] [Google Scholar]
  9. Kozak M. Adherence to the first-AUG rule when a second AUG codon follows closely upon the first. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2662–2666. doi: 10.1073/pnas.92.7.2662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Leonard J., Peers B., Johnson T., Ferreri K., Lee S., Montminy M. R. Characterization of somatostatin transactivating factor-1, a novel homeobox factor that stimulates somatostatin expression in pancreatic islet cells. Mol Endocrinol. 1993 Oct;7(10):1275–1283. doi: 10.1210/mend.7.10.7505393. [DOI] [PubMed] [Google Scholar]
  12. Lu M., Miller C., Habener J. F. Functional regions of the homeodomain protein IDX-1 required for transactivation of the rat somatostatin gene. Endocrinology. 1996 Jul;137(7):2959–2967. doi: 10.1210/endo.137.7.8770920. [DOI] [PubMed] [Google Scholar]
  13. MacFarlane W. M., Read M. L., Gilligan M., Bujalska I., Docherty K. Glucose modulates the binding activity of the beta-cell transcription factor IUF1 in a phosphorylation-dependent manner. Biochem J. 1994 Oct 15;303(Pt 2):625–631. doi: 10.1042/bj3030625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Marshak S., Totary H., Cerasi E., Melloul D. Purification of the beta-cell glucose-sensitive factor that transactivates the insulin gene differentially in normal and transformed islet cells. Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15057–15062. doi: 10.1073/pnas.93.26.15057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Melloul D., Ben-Neriah Y., Cerasi E. Glucose modulates the binding of an islet-specific factor to a conserved sequence within the rat I and the human insulin promoters. Proc Natl Acad Sci U S A. 1993 May 1;90(9):3865–3869. doi: 10.1073/pnas.90.9.3865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller C. P., McGehee R. E., Jr, Habener J. F. IDX-1: a new homeodomain transcription factor expressed in rat pancreatic islets and duodenum that transactivates the somatostatin gene. EMBO J. 1994 Mar 1;13(5):1145–1156. doi: 10.1002/j.1460-2075.1994.tb06363.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Peers B., Leonard J., Sharma S., Teitelman G., Montminy M. R. Insulin expression in pancreatic islet cells relies on cooperative interactions between the helix loop helix factor E47 and the homeobox factor STF-1. Mol Endocrinol. 1994 Dec;8(12):1798–1806. doi: 10.1210/mend.8.12.7708065. [DOI] [PubMed] [Google Scholar]
  18. Perfetti R., Rafizadeh C. M., Liotta A. S., Egan J. M. Age-dependent reduction in insulin secretion and insulin mRNA in isolated islets from rats. Am J Physiol. 1995 Dec;269(6 Pt 1):E983–E990. doi: 10.1152/ajpendo.1995.269.6.E983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Radovick S., Nations M., Du Y., Berg L. A., Weintraub B. D., Wondisford F. E. A mutation in the POU-homeodomain of Pit-1 responsible for combined pituitary hormone deficiency. Science. 1992 Aug 21;257(5073):1115–1118. doi: 10.1126/science.257.5073.1115. [DOI] [PubMed] [Google Scholar]
  20. Sharma S., Jhala U. S., Johnson T., Ferreri K., Leonard J., Montminy M. Hormonal regulation of an islet-specific enhancer in the pancreatic homeobox gene STF-1. Mol Cell Biol. 1997 May;17(5):2598–2604. doi: 10.1128/mcb.17.5.2598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stoffers D. A., Ferrer J., Clarke W. L., Habener J. F. Early-onset type-II diabetes mellitus (MODY4) linked to IPF1. Nat Genet. 1997 Oct;17(2):138–139. doi: 10.1038/ng1097-138. [DOI] [PubMed] [Google Scholar]
  22. Stoffers D. A., Zinkin N. T., Stanojevic V., Clarke W. L., Habener J. F. Pancreatic agenesis attributable to a single nucleotide deletion in the human IPF1 gene coding sequence. Nat Genet. 1997 Jan;15(1):106–110. doi: 10.1038/ng0197-106. [DOI] [PubMed] [Google Scholar]
  23. Tattersall R. B., Fajans S. S. A difference between the inheritance of classical juvenile-onset and maturity-onset type diabetes of young people. Diabetes. 1975 Jan;24(1):44–53. doi: 10.2337/diab.24.1.44. [DOI] [PubMed] [Google Scholar]
  24. Vallejo M., Penchuk L., Habener J. F. Somatostatin gene upstream enhancer element activated by a protein complex consisting of CREB, Isl-1-like, and alpha-CBF-like transcription factors. J Biol Chem. 1992 Jun 25;267(18):12876–12884. [PubMed] [Google Scholar]
  25. Waeber G., Thompson N., Nicod P., Bonny C. Transcriptional activation of the GLUT2 gene by the IPF-1/STF-1/IDX-1 homeobox factor. Mol Endocrinol. 1996 Nov;10(11):1327–1334. doi: 10.1210/mend.10.11.8923459. [DOI] [PubMed] [Google Scholar]
  26. Walker W. H., Girardet C., Habener J. F. Alternative exon splicing controls a translational switch from activator to repressor isoforms of transcription factor CREB during spermatogenesis. J Biol Chem. 1996 Aug 16;271(33):20145–21050. [PubMed] [Google Scholar]
  27. Wang Y., Perfetti R., Greig N. H., Holloway H. W., DeOre K. A., Montrose-Rafizadeh C., Elahi D., Egan J. M. Glucagon-like peptide-1 can reverse the age-related decline in glucose tolerance in rats. J Clin Invest. 1997 Jun 15;99(12):2883–2889. doi: 10.1172/JCI119482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Watada H., Kajimoto Y., Umayahara Y., Matsuoka T., Kaneto H., Fujitani Y., Kamada T., Kawamori R., Yamasaki Y. The human glucokinase gene beta-cell-type promoter: an essential role of insulin promoter factor 1/PDX-1 in its activation in HIT-T15 cells. Diabetes. 1996 Nov;45(11):1478–1488. doi: 10.2337/diab.45.11.1478. [DOI] [PubMed] [Google Scholar]
  29. Yamagata K., Furuta H., Oda N., Kaisaki P. J., Menzel S., Cox N. J., Fajans S. S., Signorini S., Stoffel M., Bell G. I. Mutations in the hepatocyte nuclear factor-4alpha gene in maturity-onset diabetes of the young (MODY1) Nature. 1996 Dec 5;384(6608):458–460. doi: 10.1038/384458a0. [DOI] [PubMed] [Google Scholar]
  30. Yamagata K., Oda N., Kaisaki P. J., Menzel S., Furuta H., Vaxillaire M., Southam L., Cox R. D., Lathrop G. M., Boriraj V. V. Mutations in the hepatocyte nuclear factor-1alpha gene in maturity-onset diabetes of the young (MODY3) Nature. 1996 Dec 5;384(6608):455–458. doi: 10.1038/384455a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES