Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1991 Sep 25;19(18):4925–4930. doi: 10.1093/nar/19.18.4925

Regulatory regions of rat insulin I gene necessary for expression in transgenic mice.

F Dandoy-Dron 1, E Monthioux 1, J Jami 1, D Bucchini 1
PMCID: PMC328791  PMID: 1923760

Abstract

Ten transgenic mouse lines harboring the -346/-103 fragment of the rat insulin I enhancer linked to a heterologous promoter and a reporter gene (Eins-Ptk-CAT construct) were produced. Expression of the hybrid transgene was essentially observed in pancreas and to a lesser extent in brain. These results indicate that the rat insulin I promoter is dispensable for pancreatic expression. This insulin gene sequence is the shortest fragment described as conferring tissue-specific expression in transgenic mice. Two short homologous sequences in the rat insulin I enhancer fragment used, IEB2 and IEB1, have been described as playing a dominant role in the regulation of HIT hamster insulinoma cell-specific transcription of the insulin gene (1). We investigated whether the combination of IEB2 and IEB1 sequences is sufficient to confer specific expression in transgenic mice to a IEB2-IEB1-Ptk-CAT gene construct. No CAT activity was observed neither in pancreas nor in any other organ examined in 19 different transgenic mice. Moreover in transient expression experiments in RIN2A rat insulinoma cells, the IEB sequences had a very weak or no enhancer activity. These observations contribute to the conclusion that DNA regulatory elements other than the IEB sequences are necessary for gene expression in vivo.

Full text

PDF
4926

Images in this article

Selected References

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

  1. Alpert S., Hanahan D., Teitelman G. Hybrid insulin genes reveal a developmental lineage for pancreatic endocrine cells and imply a relationship with neurons. Cell. 1988 Apr 22;53(2):295–308. doi: 10.1016/0092-8674(88)90391-1. [DOI] [PubMed] [Google Scholar]
  2. Begemann M., Tan S. S., Cunningham B. A., Edelman G. M. Expression of chicken liver cell adhesion molecule fusion genes in transgenic mice. Proc Natl Acad Sci U S A. 1990 Nov;87(22):9042–9046. doi: 10.1073/pnas.87.22.9042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boam D. S., Clark A. R., Docherty K. Positive and negative regulation of the human insulin gene by multiple trans-acting factors. J Biol Chem. 1990 May 15;265(14):8285–8296. [PubMed] [Google Scholar]
  4. Boam D. S., Docherty K. A tissue-specific nuclear factor binds to multiple sites in the human insulin-gene enhancer. Biochem J. 1989 Nov 15;264(1):233–239. doi: 10.1042/bj2640233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cordell B., Bell G., Tischer E., DeNoto F. M., Ullrich A., Pictet R., Rutter W. J., Goodman H. M. Isolation and characterization of a cloned rat insulin gene. Cell. 1979 Oct;18(2):533–543. doi: 10.1016/0092-8674(79)90070-9. [DOI] [PubMed] [Google Scholar]
  6. Crowe D. T., Tsai M. J. Mutagenesis of the rat insulin II 5'-flanking region defines sequences important for expression in HIT cells. Mol Cell Biol. 1989 Apr;9(4):1784–1789. doi: 10.1128/mcb.9.4.1784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Edlund T., Walker M. D., Barr P. J., Rutter W. J. Cell-specific expression of the rat insulin gene: evidence for role of two distinct 5' flanking elements. Science. 1985 Nov 22;230(4728):912–916. doi: 10.1126/science.3904002. [DOI] [PubMed] [Google Scholar]
  8. Fiedorek F. T., Jr, Carnaghi L. R., Giddings S. J. Selective expression of the insulin I gene in rat insulinoma-derived cell lines. Mol Endocrinol. 1990 Jul;4(7):990–999. doi: 10.1210/mend-4-7-990. [DOI] [PubMed] [Google Scholar]
  9. Fromont-Racine M., Bucchini D., Madsen O., Desbois P., Linde S., Nielsen J. H., Saulnier C., Ripoche M. A., Jami J., Pictet R. Effect of 5'-flanking sequence deletions on expression of the human insulin gene in transgenic mice. Mol Endocrinol. 1990 May;4(5):669–677. doi: 10.1210/mend-4-5-669. [DOI] [PubMed] [Google Scholar]
  10. Gazdar A. F., Chick W. L., Oie H. K., Sims H. L., King D. L., Weir G. C., Lauris V. Continuous, clonal, insulin- and somatostatin-secreting cell lines established from a transplantable rat islet cell tumor. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3519–3523. doi: 10.1073/pnas.77.6.3519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  13. Hammer R. E., Swift G. H., Ornitz D. M., Quaife C. J., Palmiter R. D., Brinster R. L., MacDonald R. J. The rat elastase I regulatory element is an enhancer that directs correct cell specificity and developmental onset of expression in transgenic mice. Mol Cell Biol. 1987 Aug;7(8):2956–2967. doi: 10.1128/mcb.7.8.2956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hanahan D. Heritable formation of pancreatic beta-cell tumours in transgenic mice expressing recombinant insulin/simian virus 40 oncogenes. Nature. 1985 May 9;315(6015):115–122. doi: 10.1038/315115a0. [DOI] [PubMed] [Google Scholar]
  15. Hwung Y. P., Gu Y. Z., Tsai M. J. Cooperativity of sequence elements mediates tissue specificity of the rat insulin II gene. Mol Cell Biol. 1990 Apr;10(4):1784–1788. doi: 10.1128/mcb.10.4.1784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Karlsson O., Edlund T., Moss J. B., Rutter W. J., Walker M. D. A mutational analysis of the insulin gene transcription control region: expression in beta cells is dependent on two related sequences within the enhancer. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8819–8823. doi: 10.1073/pnas.84.24.8819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Karlsson O., Thor S., Norberg T., Ohlsson H., Edlund T. Insulin gene enhancer binding protein Isl-1 is a member of a novel class of proteins containing both a homeo- and a Cys-His domain. Nature. 1990 Apr 26;344(6269):879–882. doi: 10.1038/344879a0. [DOI] [PubMed] [Google Scholar]
  18. Karlsson O., Walker M. D., Rutter W. J., Edlund T. Individual protein-binding domains of the insulin gene enhancer positively activate beta-cell-specific transcription. Mol Cell Biol. 1989 Feb;9(2):823–827. doi: 10.1128/mcb.9.2.823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lee K. F., Atiee S. H., Rosen J. M. Differential regulation of rat beta-casein-chloramphenicol acetyltransferase fusion gene expression in transgenic mice. Mol Cell Biol. 1989 Feb;9(2):560–565. doi: 10.1128/mcb.9.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lomedico P., Rosenthal N., Efstratidadis A., Gilbert W., Kolodner R., Tizard R. The structure and evolution of the two nonallelic rat preproinsulin genes. Cell. 1979 Oct;18(2):545–558. doi: 10.1016/0092-8674(79)90071-0. [DOI] [PubMed] [Google Scholar]
  21. Nguyen V. T., Morange M., Bensaude O. Firefly luciferase luminescence assays using scintillation counters for quantitation in transfected mammalian cells. Anal Biochem. 1988 Jun;171(2):404–408. doi: 10.1016/0003-2697(88)90505-2. [DOI] [PubMed] [Google Scholar]
  22. Ohlsson H., Karlsson O., Edlund T. A beta-cell-specific protein binds to the two major regulatory sequences of the insulin gene enhancer. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4228–4231. doi: 10.1073/pnas.85.12.4228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sarvetnick N., Liggitt D., Pitts S. L., Hansen S. E., Stewart T. A. Insulin-dependent diabetes mellitus induced in transgenic mice by ectopic expression of class II MHC and interferon-gamma. Cell. 1988 Mar 11;52(5):773–782. doi: 10.1016/0092-8674(88)90414-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shibasaki Y., Sakura H., Takaku F., Kasuga M. Insulin enhancer binding protein has helix-loop-helix structure. Biochem Biophys Res Commun. 1990 Jul 16;170(1):314–321. doi: 10.1016/0006-291x(90)91276-x. [DOI] [PubMed] [Google Scholar]
  25. Soares M. B., Schon E., Henderson A., Karathanasis S. K., Cate R., Zeitlin S., Chirgwin J., Efstratiadis A. RNA-mediated gene duplication: the rat preproinsulin I gene is a functional retroposon. Mol Cell Biol. 1985 Aug;5(8):2090–2103. doi: 10.1128/mcb.5.8.2090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Teitelman G., Lee J. K. Cell lineage analysis of pancreatic islet development: glucagon and insulin cells arise from catecholaminergic precursors present in the pancreatic duct. Dev Biol. 1987 Jun;121(2):454–466. doi: 10.1016/0012-1606(87)90182-5. [DOI] [PubMed] [Google Scholar]
  27. Walker M. D., Edlund T., Boulet A. M., Rutter W. J. Cell-specific expression controlled by the 5'-flanking region of insulin and chymotrypsin genes. Nature. 1983 Dec 8;306(5943):557–561. doi: 10.1038/306557a0. [DOI] [PubMed] [Google Scholar]
  28. Walker M. D., Park C. W., Rosen A., Aronheim A. A cDNA from a mouse pancreatic beta cell encoding a putative transcription factor of the insulin gene. Nucleic Acids Res. 1990 Mar 11;18(5):1159–1166. doi: 10.1093/nar/18.5.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Whelan J., Cordle S. R., Henderson E., Weil P. A., Stein R. Identification of a pancreatic beta-cell insulin gene transcription factor that binds to and appears to activate cell-type-specific expression: its possible relationship to other cellular factors that bind to a common insulin gene sequence. Mol Cell Biol. 1990 Apr;10(4):1564–1572. doi: 10.1128/mcb.10.4.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Whelan J., Poon D., Weil P. A., Stein R. Pancreatic beta-cell-type-specific expression of the rat insulin II gene is controlled by positive and negative cellular transcriptional elements. Mol Cell Biol. 1989 Aug;9(8):3253–3259. doi: 10.1128/mcb.9.8.3253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. de Wet J. R., Wood K. V., DeLuca M., Helinski D. R., Subramani S. Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol. 1987 Feb;7(2):725–737. doi: 10.1128/mcb.7.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES