Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Dec;84(24):8946–8950. doi: 10.1073/pnas.84.24.8946

Differential expression of alternative 5' untranslated regions in mRNAs encoding rat insulin-like growth factor I.

W L Lowe Jr 1, C T Roberts Jr 1, S R Lasky 1, D LeRoith 1
PMCID: PMC299668  PMID: 3480521

Abstract

Rat insulin-like growth factor I (IGF-I) cDNAs contain three alternative 5' untranslated sequences (termed class A, B, and C), which are associated with an identical coding region for the mature IGF-I peptide. A solution hybridization/RNase protection assay was used to simultaneously quantitate the relative abundance of IGF-I transcripts with the different 5' untranslated regions. In all the tissues studied, transcripts with the class C 5' untranslated region were most abundant. In contrast, both class A and B transcripts were tissue specific. Class A transcripts were present in moderate abundance in liver; in low abundance in kidney, lung, testes, and stomach; and were undetectable in muscle, heart, and brain; whereas class B transcripts were detected only in liver. These three classes of 5' untranslated region were also regulated independently by growth hormone. In liver, heart, kidney, and lung, growth hormone increased the abundance of class C transcripts 2- to 3-fold. In liver, growth hormone increased the abundance of the class A and B transcripts 6- to 7-fold. In lung and kidney, on the other hand, the abundance of class A transcripts was not affected by growth hormone. Thus, rat IGF-I gene transcripts contain one of three alternative 5' untranslated regions, which are expressed in a tissue-specific manner and are differentially regulated by growth hormone. Finally, cDNA probes unique to two of the three 5' untranslated regions hybridized to all three major species of IGF-I mRNA typically seen on RNA blots with a coding region probe.

Full text

PDF
8946

Images in this article

8948Image
on p.8948
8949Image
on p.8949
8950Image
on p.8950
8950Image
on p.8950
8950Image
on p.8950

Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell G. I., Stempien M. M., Fong N. M., Rall L. B. Sequences of liver cDNAs encoding two different mouse insulin-like growth factor I precursors. Nucleic Acids Res. 1986 Oct 24;14(20):7873–7882. doi: 10.1093/nar/14.20.7873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benyajati C., Spoerel N., Haymerle H., Ashburner M. The messenger RNA for alcohol dehydrogenase in Drosophila melanogaster differs in its 5' end in different developmental stages. Cell. 1983 May;33(1):125–133. doi: 10.1016/0092-8674(83)90341-0. [DOI] [PubMed] [Google Scholar]
  4. Bond B. J., Davidson N. The Drosophila melanogaster actin 5C gene uses two transcription initiation sites and three polyadenylation sites to express multiple mRNA species. Mol Cell Biol. 1986 Jun;6(6):2080–2088. doi: 10.1128/mcb.6.6.2080. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brawerman G. Determinants of messenger RNA stability. Cell. 1987 Jan 16;48(1):5–6. doi: 10.1016/0092-8674(87)90346-1. [DOI] [PubMed] [Google Scholar]
  6. Brissenden J. E., Ullrich A., Francke U. Human chromosomal mapping of genes for insulin-like growth factors I and II and epidermal growth factor. 1984 Aug 30-Sep 5Nature. 310(5980):781–784. doi: 10.1038/310781a0. [DOI] [PubMed] [Google Scholar]
  7. Cathala G., Savouret J. F., Mendez B., West B. L., Karin M., Martial J. A., Baxter J. D. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. doi: 10.1089/dna.1983.2.329. [DOI] [PubMed] [Google Scholar]
  8. D'Ercole A. J., Stiles A. D., Underwood L. E. Tissue concentrations of somatomedin C: further evidence for multiple sites of synthesis and paracrine or autocrine mechanisms of action. Proc Natl Acad Sci U S A. 1984 Feb;81(3):935–939. doi: 10.1073/pnas.81.3.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  10. Frunzio R., Chiariotti L., Brown A. L., Graham D. E., Rechler M. M., Bruni C. B. Structure and expression of the rat insulin-like growth factor II (rIGF-II) gene. rIGF-II RNAs are transcribed from two promoters. J Biol Chem. 1986 Dec 25;261(36):17138–17149. [PubMed] [Google Scholar]
  11. Han V. K., D'Ercole A. J., Lund P. K. Cellular localization of somatomedin (insulin-like growth factor) messenger RNA in the human fetus. Science. 1987 Apr 10;236(4798):193–197. doi: 10.1126/science.3563497. [DOI] [PubMed] [Google Scholar]
  12. Hynes M. A., Van Wyk J. J., Brooks P. J., D'Ercole A. J., Jansen M., Lund P. K. Growth hormone dependence of somatomedin-C/insulin-like growth factor-I and insulin-like growth factor-II messenger ribonucleic acids. Mol Endocrinol. 1987 Mar;1(3):233–242. doi: 10.1210/mend-1-3-233. [DOI] [PubMed] [Google Scholar]
  13. Jansen M., van Schaik F. M., Ricker A. T., Bullock B., Woods D. E., Gabbay K. H., Nussbaum A. L., Sussenbach J. S., Van den Brande J. L. Sequence of cDNA encoding human insulin-like growth factor I precursor. Nature. 1983 Dec 8;306(5943):609–611. doi: 10.1038/306609a0. [DOI] [PubMed] [Google Scholar]
  14. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kozak M. Influences of mRNA secondary structure on initiation by eukaryotic ribosomes. Proc Natl Acad Sci U S A. 1986 May;83(9):2850–2854. doi: 10.1073/pnas.83.9.2850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leff S. E., Rosenfeld M. G., Evans R. M. Complex transcriptional units: diversity in gene expression by alternative RNA processing. Annu Rev Biochem. 1986;55:1091–1117. doi: 10.1146/annurev.bi.55.070186.005303. [DOI] [PubMed] [Google Scholar]
  17. Lowe W. L., Jr, Schaffner A. E., Roberts C. T., Jr, LeRoith D. Developmental regulation of somatostatin gene expression in the brain is region specific. Mol Endocrinol. 1987 Feb;1(2):181–187. doi: 10.1210/mend-1-2-181. [DOI] [PubMed] [Google Scholar]
  18. Lund P. K., Moats-Staats B. M., Hynes M. A., Simmons J. G., Jansen M., D'Ercole A. J., Van Wyk J. J. Somatomedin-C/insulin-like growth factor-I and insulin-like growth factor-II mRNAs in rat fetal and adult tissues. J Biol Chem. 1986 Nov 5;261(31):14539–14544. [PubMed] [Google Scholar]
  19. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Murphy L. J., Bell G. I., Friesen H. G. Growth hormone stimulates sequential induction of c-myc and insulin-like growth factor I expression in vivo. Endocrinology. 1987 May;120(5):1806–1812. doi: 10.1210/endo-120-5-1806. [DOI] [PubMed] [Google Scholar]
  21. Murphy L. J., Bell G. I., Friesen H. G. Tissue distribution of insulin-like growth factor I and II messenger ribonucleic acid in the adult rat. Endocrinology. 1987 Apr;120(4):1279–1282. doi: 10.1210/endo-120-4-1279. [DOI] [PubMed] [Google Scholar]
  22. Pelletier J., Sonenberg N. Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency. Cell. 1985 Mar;40(3):515–526. doi: 10.1016/0092-8674(85)90200-4. [DOI] [PubMed] [Google Scholar]
  23. Roberts C. T., Jr, Brown A. L., Graham D. E., Seelig S., Berry S., Gabbay K. H., Rechler M. M. Growth hormone regulates the abundance of insulin-like growth factor I RNA in adult rat liver. J Biol Chem. 1986 Aug 5;261(22):10025–10028. [PubMed] [Google Scholar]
  24. Roberts C. T., Jr, Lasky S. R., Lowe W. L., Jr, LeRoith D. Rat IGF-I cDNA's contain multiple 5'-untranslated regions. Biochem Biophys Res Commun. 1987 Aug 14;146(3):1154–1159. doi: 10.1016/0006-291x(87)90768-6. [DOI] [PubMed] [Google Scholar]
  25. Roberts C. T., Jr, Lasky S. R., Lowe W. L., Jr, Seaman W. T., LeRoith D. Molecular cloning of rat insulin-like growth factor I complementary deoxyribonucleic acids: differential messenger ribonucleic acid processing and regulation by growth hormone in extrahepatic tissues. Mol Endocrinol. 1987 Mar;1(3):243–248. doi: 10.1210/mend-1-3-243. [DOI] [PubMed] [Google Scholar]
  26. Rotwein P. Two insulin-like growth factor I messenger RNAs are expressed in human liver. Proc Natl Acad Sci U S A. 1986 Jan;83(1):77–81. doi: 10.1073/pnas.83.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schibler U., Hagenbüchle O., Wellauer P. K., Pittet A. C. Two promoters of different strengths control the transcription of the mouse alpha-amylase gene Amy-1a in the parotid gland and the liver. Cell. 1983 Jun;33(2):501–508. doi: 10.1016/0092-8674(83)90431-2. [DOI] [PubMed] [Google Scholar]
  28. Shaw P., Sordat B., Schibler U. The two promoters of the mouse alpha-amylase gene Amy-1a are differentially activated during parotid gland differentiation. Cell. 1985 Apr;40(4):907–912. doi: 10.1016/0092-8674(85)90350-2. [DOI] [PubMed] [Google Scholar]
  29. Shimatsu A., Rotwein P. Mosaic evolution of the insulin-like growth factors. Organization, sequence, and expression of the rat insulin-like growth factor I gene. J Biol Chem. 1987 Jun 5;262(16):7894–7900. [PubMed] [Google Scholar]
  30. Soares M. B., Turken A., Ishii D., Mills L., Episkopou V., Cotter S., Zeitlin S., Efstratiadis A. Rat insulin-like growth factor II gene. A single gene with two promoters expressing a multitranscript family. J Mol Biol. 1986 Dec 20;192(4):737–752. doi: 10.1016/0022-2836(86)90025-2. [DOI] [PubMed] [Google Scholar]
  31. Spena A., Krause E., Dobberstein B. Translation efficiency of zein mRNA is reduced by hybrid formation between the 5'- and 3'-untranslated region. EMBO J. 1985 Sep;4(9):2153–2158. doi: 10.1002/j.1460-2075.1985.tb03909.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Starksen N. F., Simpson P. C., Bishopric N., Coughlin S. R., Lee W. M., Escobedo J. A., Williams L. T. Cardiac myocyte hypertrophy is associated with c-myc protooncogene expression. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8348–8350. doi: 10.1073/pnas.83.21.8348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Young R. A., Hagenbüchle O., Schibler U. A single mouse alpha-amylase gene specifies two different tissue-specific mRNAs. Cell. 1981 Feb;23(2):451–458. doi: 10.1016/0092-8674(81)90140-9. [DOI] [PubMed] [Google Scholar]
  34. de Pagter-Holthuizen P., Jansen M., van Schaik F. M., van der Kammen R., Oosterwijk C., Van den Brande J. L., Sussenbach J. S. The human insulin-like growth factor II gene contains two development-specific promoters. FEBS Lett. 1987 Apr 20;214(2):259–264. doi: 10.1016/0014-5793(87)80066-2. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES