Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1987 Sep;6(9):2565–2572. doi: 10.1002/j.1460-2075.1987.tb02545.x

Organization of the fibronectin gene provides evidence for exon shuffling during evolution.

R S Patel 1, E Odermatt 1, J E Schwarzbauer 1, R O Hynes 1
PMCID: PMC553675  PMID: 3119323

Abstract

We report the organization of the two ends of the rat fibronectin gene which encode the type I and II repeating units of the protein. We show that each of these modular structural units is encoded by a separate exon. Homologous type I and II repeats are known to occur in tissue plasminogen activator, factor XII and a bovine seminal plasma protein. Comparison of these sequences and the exon structures of the fibronectin and tissue plasminogen activator genes indicates that exons encoding type I and type II repeats have reassorted during evolution. We also report analyses of the extreme 5' and 3' ends of the fibronectin gene including the promoter region and the exon encoding the prepro sequence of fibronectin and we show that the gene is transcribed from a single initiation site to a single polyadenylation site. These data provide information pertinent to the transcriptional regulation of the gene, the alternative splicing of the primary transcript and the structure of the primary translation product.

Full text

PDF
2565

Images in this article

2570Fig. 6.
on p.2570

Selected References

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

  1. Adams S. L., Boettiger D., Focht R. J., Holtzer H., Pacifici M. Regulation of the synthesis of extracellular matrix components in chondroblasts transformed by a temperature-sensitive mutant of Rous sarcoma virus. Cell. 1982 Sep;30(2):373–384. doi: 10.1016/0092-8674(82)90235-5. [DOI] [PubMed] [Google Scholar]
  2. Allebach E. S., Boettiger D., Pacifici M., Adams S. L. Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation. Mol Cell Biol. 1985 May;5(5):1002–1008. doi: 10.1128/mcb.5.5.1002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker M. E. The PDC-109 protein from bovine seminal plasma is similar to the gelatin-binding domain of bovine fibronectin and a kringle domain of human tissue-type plasminogen activator. Biochem Biophys Res Commun. 1985 Aug 15;130(3):1010–1014. doi: 10.1016/0006-291x(85)91715-2. [DOI] [PubMed] [Google Scholar]
  4. Bányai L., Váradi A., Patthy L. Common evolutionary origin of the fibrin-binding structures of fibronectin and tissue-type plasminogen activator. FEBS Lett. 1983 Oct 31;163(1):37–41. doi: 10.1016/0014-5793(83)81157-0. [DOI] [PubMed] [Google Scholar]
  5. Caput D., Beutler B., Hartog K., Thayer R., Brown-Shimer S., Cerami A. Identification of a common nucleotide sequence in the 3'-untranslated region of mRNA molecules specifying inflammatory mediators. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1670–1674. doi: 10.1073/pnas.83.6.1670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Dente L., Ciliberto G., Cortese R. Structure of the human alpha 1-acid glycoprotein gene: sequence homology with other human acute phase protein genes. Nucleic Acids Res. 1985 Jun 11;13(11):3941–3952. doi: 10.1093/nar/13.11.3941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dynan W. S., Tjian R. Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins. 1985 Aug 29-Sep 4Nature. 316(6031):774–778. doi: 10.1038/316774a0. [DOI] [PubMed] [Google Scholar]
  9. Ebright R. H., Cossart P., Gicquel-Sanzey B., Beckwith J. Mutations that alter the DNA sequence specificity of the catabolite gene activator protein of E. coli. Nature. 1984 Sep 20;311(5983):232–235. doi: 10.1038/311232a0. [DOI] [PubMed] [Google Scholar]
  10. Esch F. S., Ling N. C., Böhlen P., Ying S. Y., Guillemin R. Primary structure of PDC-109, a major protein constituent of bovine seminal plasma. Biochem Biophys Res Commun. 1983 Jun 29;113(3):861–867. doi: 10.1016/0006-291x(83)91078-1. [DOI] [PubMed] [Google Scholar]
  11. Friedman R. L., Stark G. R. alpha-Interferon-induced transcription of HLA and metallothionein genes containing homologous upstream sequences. Nature. 1985 Apr 18;314(6012):637–639. doi: 10.1038/314637a0. [DOI] [PubMed] [Google Scholar]
  12. Gilbert W. Why genes in pieces? Nature. 1978 Feb 9;271(5645):501–501. doi: 10.1038/271501a0. [DOI] [PubMed] [Google Scholar]
  13. Gutman A., Yamada K. M., Kornblihtt A. Human fibronectin is synthesized as a pre-propolypeptide. FEBS Lett. 1986 Oct 20;207(1):145–148. doi: 10.1016/0014-5793(86)80029-1. [DOI] [PubMed] [Google Scholar]
  14. Hirano H., Yamada Y., Sullivan M., de Crombrugghe B., Pastan I., Yamada K. M. Isolation of genomic DNA clones spanning the entire fibronectin gene. Proc Natl Acad Sci U S A. 1983 Jan;80(1):46–50. doi: 10.1073/pnas.80.1.46. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hynes R. O. Fibronectins. Sci Am. 1986 Jun;254(6):42–51. doi: 10.1038/scientificamerican0686-42. [DOI] [PubMed] [Google Scholar]
  16. Hynes R. O., Yamada K. M. Fibronectins: multifunctional modular glycoproteins. J Cell Biol. 1982 Nov;95(2 Pt 1):369–377. doi: 10.1083/jcb.95.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hynes R. Molecular biology of fibronectin. Annu Rev Cell Biol. 1985;1:67–90. doi: 10.1146/annurev.cb.01.110185.000435. [DOI] [PubMed] [Google Scholar]
  18. Jochemsen A. G., Bernards R., van Kranen H. J., Houweling A., Bos J. L., van der Eb A. J. Different activities of the adenovirus types 5 and 12 E1A regions in transformation with the EJ Ha-ras oncogene. J Virol. 1986 Sep;59(3):684–691. doi: 10.1128/jvi.59.3.684-691.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Karin M., Haslinger A., Holtgreve H., Richards R. I., Krauter P., Westphal H. M., Beato M. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature. 1984 Apr 5;308(5959):513–519. doi: 10.1038/308513a0. [DOI] [PubMed] [Google Scholar]
  20. Korman A. J., Frantz J. D., Strominger J. L., Mulligan R. C. Expression of human class II major histocompatibility complex antigens using retrovirus vectors. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2150–2154. doi: 10.1073/pnas.84.8.2150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kornblihtt A. R., Umezawa K., Vibe-Pedersen K., Baralle F. E. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J. 1985 Jul;4(7):1755–1759. doi: 10.1002/j.1460-2075.1985.tb03847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kornblihtt A. R., Vibe-Pedersen K., Baralle F. E. Isolation and characterization of cDNA clones for human and bovine fibronectins. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3218–3222. doi: 10.1073/pnas.80.11.3218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Leibovitch S. A., Hillion J., Leibovitch M. P., Guillier M., Schmitz A., Harel J. Expression of extracellular matrix genes in relation to myogenesis and neoplastic transformation. Exp Cell Res. 1986 Oct;166(2):526–534. doi: 10.1016/0014-4827(86)90497-0. [DOI] [PubMed] [Google Scholar]
  25. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  26. McMullen B. A., Fujikawa K. Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). J Biol Chem. 1985 May 10;260(9):5328–5341. [PubMed] [Google Scholar]
  27. Montminy M. R., Sevarino K. A., Wagner J. A., Mandel G., Goodman R. H. Identification of a cyclic-AMP-responsive element within the rat somatostatin gene. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6682–6686. doi: 10.1073/pnas.83.18.6682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moore D. D., Marks A. R., Buckley D. I., Kapler G., Payvar F., Goodman H. M. The first intron of the human growth hormone gene contains a binding site for glucocorticoid receptor. Proc Natl Acad Sci U S A. 1985 Feb;82(3):699–702. doi: 10.1073/pnas.82.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ny T., Elgh F., Lund B. The structure of the human tissue-type plasminogen activator gene: correlation of intron and exon structures to functional and structural domains. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5355–5359. doi: 10.1073/pnas.81.17.5355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Odermatt E., Tamkun J. W., Hynes R. O. Repeating modular structure of the fibronectin gene: relationship to protein structure and subunit variation. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6571–6575. doi: 10.1073/pnas.82.19.6571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Oldberg A., Ruoslahti E. Evolution of the fibronectin gene. Exon structure of cell attachment domain. J Biol Chem. 1986 Feb 15;261(5):2113–2116. [PubMed] [Google Scholar]
  32. Oliver N., Newby R. F., Furcht L. T., Bourgeois S. Regulation of fibronectin biosynthesis by glucocorticoids in human fibrosarcoma cells and normal fibroblasts. Cell. 1983 May;33(1):287–296. doi: 10.1016/0092-8674(83)90357-4. [DOI] [PubMed] [Google Scholar]
  33. Owens R. J., Baralle F. E. Exon structure of the collagen-binding domain of human fibronectin. FEBS Lett. 1986 Aug 18;204(2):318–322. doi: 10.1016/0014-5793(86)80836-5. [DOI] [PubMed] [Google Scholar]
  34. Payvar F., DeFranco D., Firestone G. L., Edgar B., Wrange O., Okret S., Gustafsson J. A., Yamamoto K. R. Sequence-specific binding of glucocorticoid receptor to MTV DNA at sites within and upstream of the transcribed region. Cell. 1983 Dec;35(2 Pt 1):381–392. doi: 10.1016/0092-8674(83)90171-x. [DOI] [PubMed] [Google Scholar]
  35. Pick-Kober K. H., Münker D., Gressner A. M. Fibronectin is synthesized as an acute phase reactant in rat hepatocytes. J Clin Chem Clin Biochem. 1986 Aug;24(8):521–528. doi: 10.1515/cclm.1986.24.8.521. [DOI] [PubMed] [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schwarzbauer J. E., Mulligan R. C., Hynes R. O. Efficient and stable expression of recombinant fibronectin polypeptides. Proc Natl Acad Sci U S A. 1987 Feb;84(3):754–758. doi: 10.1073/pnas.84.3.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schwarzbauer J. E., Patel R. S., Fonda D., Hynes R. O. Multiple sites of alternative splicing of the rat fibronectin gene transcript. EMBO J. 1987 Sep;6(9):2573–2580. doi: 10.1002/j.1460-2075.1987.tb02547.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schwarzbauer J. E., Tamkun J. W., Lemischka I. R., Hynes R. O. Three different fibronectin mRNAs arise by alternative splicing within the coding region. Cell. 1983 Dec;35(2 Pt 1):421–431. doi: 10.1016/0092-8674(83)90175-7. [DOI] [PubMed] [Google Scholar]
  40. Senger D. R., Destree A. T., Hynes R. O. Complex regulation of fibronectin synthesis by cells in culture. Am J Physiol. 1983 Jul;245(1):C144–C150. doi: 10.1152/ajpcell.1983.245.1.C144. [DOI] [PubMed] [Google Scholar]
  41. Skorstengaard K., Jensen M. S., Sahl P., Petersen T. E., Magnusson S. Complete primary structure of bovine plasma fibronectin. Eur J Biochem. 1986 Dec 1;161(2):441–453. doi: 10.1111/j.1432-1033.1986.tb10464.x. [DOI] [PubMed] [Google Scholar]
  42. Skorstengaard K., Thøgersen H. C., Petersen T. E. Complete primary structure of the collagen-binding domain of bovine fibronectin. Eur J Biochem. 1984 Apr 16;140(2):235–243. doi: 10.1111/j.1432-1033.1984.tb08092.x. [DOI] [PubMed] [Google Scholar]
  43. Skorstengaard K., Thøgersen H. C., Vibe-Pedersen K., Petersen T. E., Magnusson S. Purification of twelve cyanogen bromide fragments from bovine plasma fibronectin and the amino acid sequence of eight of them. Overlap evidence aligning two plasmic fragments, internal homology in gelatin-binding region and phosphorylation site near C terminus. Eur J Biochem. 1982 Nov 15;128(2-3):605–623. doi: 10.1111/j.1432-1033.1982.tb07007.x. [DOI] [PubMed] [Google Scholar]
  44. Tamkun J. W., Schwarzbauer J. E., Hynes R. O. A single rat fibronectin gene generates three different mRNAs by alternative splicing of a complex exon. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5140–5144. doi: 10.1073/pnas.81.16.5140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tyagi J. S., Hirano H., Merlino G. T., Pastan I. Transcriptional control of the fibronectin gene in chick embryo fibroblasts transformed by Rous sarcoma virus. J Biol Chem. 1983 May 10;258(9):5787–5793. [PubMed] [Google Scholar]
  46. Tyagi J. S., Hirano H., Pastan I. Modulation of fibronectin gene activity in chick embryo fibroblasts transformed by a temperature-sensitive strain (ts68) of Rous sarcoma virus. Nucleic Acids Res. 1985 Nov 25;13(22):8275–8284. doi: 10.1093/nar/13.22.8275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Vibe-Pedersen K., Kornblihtt A. R., Baralle F. E. Expression of a human alpha-globin/fibronectin gene hybrid generates two mRNAs by alternative splicing. EMBO J. 1984 Nov;3(11):2511–2516. doi: 10.1002/j.1460-2075.1984.tb02165.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Yamada K. M. Cell surface interactions with extracellular materials. Annu Rev Biochem. 1983;52:761–799. doi: 10.1146/annurev.bi.52.070183.003553. [DOI] [PubMed] [Google Scholar]
  49. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]
  51. von der Ahe D., Renoir J. M., Buchou T., Baulieu E. E., Beato M. Receptors for glucocorticosteroid and progesterone recognize distinct features of a DNA regulatory element. Proc Natl Acad Sci U S A. 1986 May;83(9):2817–2821. doi: 10.1073/pnas.83.9.2817. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES