Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1993 Dec 15;296(Pt 3):837–842. doi: 10.1042/bj2960837

Molecular cloning and tissue distribution of pig transforming growth factor alpha.

T J Vaughan 1, P S James 1, J C Pascall 1, K D Brown 1
PMCID: PMC1137770  PMID: 8280083

Abstract

Transforming growth factor alpha (TGF alpha) was originally identified as a product of tumour tissues and transformed cells in culture. Although it is now clear that expression of this factor is not restricted to neoplastic cells, there remains relatively little information about the sites of expression of TGF alpha in normal tissues. Therefore, an amplified DNA fragment encoding the pig TGF alpha precursor was cloned by reverse transcription-PCR (RT-PCR) using RNA isolated from normal skin tissue as the template. Nucleotide sequence analysis predicts a 160-residue transmembrane polypeptide that differs from the rat, mouse and human TGF alpha precursors at 14, 15 and six sites respectively. The distribution of TGF alpha mRNA in a wide variety of pig tissues was analysed by RT-PCR, using oligonucleotide primers based on the pig TGF alpha cDNA sequence. TGF alpha transcripts were detected in RNA isolated from 17 of the 22 tissues analysed, including four previously unreported sites. Using an antibody raised against a synthetic TGF alpha peptide, we have immunolocalized TGF alpha protein to cells within the red pulp of the spleen and to the distal convoluted tubules of the kidney.

Full text

PDF
837

Images in this article

Selected References

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

  1. Atkinson S., Smith P. A., Taylor J. J., Kendall-Taylor P. Specific epidermal growth factor receptors on porcine thyroid cell membranes. FEBS Lett. 1983 Mar 7;153(1):88–92. doi: 10.1016/0014-5793(83)80124-0. [DOI] [PubMed] [Google Scholar]
  2. Barrandon Y., Green H. Cell migration is essential for sustained growth of keratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor. Cell. 1987 Sep 25;50(7):1131–1137. doi: 10.1016/0092-8674(87)90179-6. [DOI] [PubMed] [Google Scholar]
  3. Barton C. M., Hall P. A., Hughes C. M., Gullick W. J., Lemoine N. R. Transforming growth factor alpha and epidermal growth factor in human pancreatic cancer. J Pathol. 1991 Feb;163(2):111–116. doi: 10.1002/path.1711630206. [DOI] [PubMed] [Google Scholar]
  4. Blasband A. J., Rogers K. T., Chen X. R., Azizkhan J. C., Lee D. C. Characterization of the rat transforming growth factor alpha gene and identification of promoter sequences. Mol Cell Biol. 1990 May;10(5):2111–2121. doi: 10.1128/mcb.10.5.2111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bosenberg M. W., Pandiella A., Massagué J. The cytoplasmic carboxy-terminal amino acid specifies cleavage of membrane TGF alpha into soluble growth factor. Cell. 1992 Dec 24;71(7):1157–1165. doi: 10.1016/s0092-8674(05)80064-9. [DOI] [PubMed] [Google Scholar]
  6. Brachmann R., Lindquist P. B., Nagashima M., Kohr W., Lipari T., Napier M., Derynck R. Transmembrane TGF-alpha precursors activate EGF/TGF-alpha receptors. Cell. 1989 Feb 24;56(4):691–700. doi: 10.1016/0092-8674(89)90591-6. [DOI] [PubMed] [Google Scholar]
  7. Bringman T. S., Lindquist P. B., Derynck R. Different transforming growth factor-alpha species are derived from a glycosylated and palmitoylated transmembrane precursor. Cell. 1987 Feb 13;48(3):429–440. doi: 10.1016/0092-8674(87)90194-2. [DOI] [PubMed] [Google Scholar]
  8. Brown P. I., Lam R., Lakshmanan J., Fisher D. A. Transforming growth factor alpha in developing rats. Am J Physiol. 1990 Aug;259(2 Pt 1):E256–E260. doi: 10.1152/ajpendo.1990.259.2.E256. [DOI] [PubMed] [Google Scholar]
  9. Brändli A. W., Adamson E. D., Simons K. Transcytosis of epidermal growth factor. The epidermal growth factor receptor mediates uptake but not transcytosis. J Biol Chem. 1991 May 5;266(13):8560–8566. [PubMed] [Google Scholar]
  10. Burgess A. W. Epidermal growth factor and transforming growth factor alpha. Br Med Bull. 1989 Apr;45(2):401–424. doi: 10.1093/oxfordjournals.bmb.a072331. [DOI] [PubMed] [Google Scholar]
  11. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  12. Coffey R. J., Jr, Derynck R., Wilcox J. N., Bringman T. S., Goustin A. S., Moses H. L., Pittelkow M. R. Production and auto-induction of transforming growth factor-alpha in human keratinocytes. 1987 Aug 27-Sep 2Nature. 328(6133):817–820. doi: 10.1038/328817a0. [DOI] [PubMed] [Google Scholar]
  13. Dagogo-Jack S., Atkinson S., Kendall-Taylor P. Demonstration of significant epidermal growth factor activity in murine thyroid tissue. J Endocrinol. 1986 Jul;110(1):67–72. doi: 10.1677/joe.0.1100067. [DOI] [PubMed] [Google Scholar]
  14. Derynck R., Goeddel D. V., Ullrich A., Gutterman J. U., Williams R. D., Bringman T. S., Berger W. H. Synthesis of messenger RNAs for transforming growth factors alpha and beta and the epidermal growth factor receptor by human tumors. Cancer Res. 1987 Feb 1;47(3):707–712. [PubMed] [Google Scholar]
  15. Derynck R., Roberts A. B., Winkler M. E., Chen E. Y., Goeddel D. V. Human transforming growth factor-alpha: precursor structure and expression in E. coli. Cell. 1984 Aug;38(1):287–297. doi: 10.1016/0092-8674(84)90550-6. [DOI] [PubMed] [Google Scholar]
  16. Elder J. T., Fisher G. J., Lindquist P. B., Bennett G. L., Pittelkow M. R., Coffey R. J., Jr, Ellingsworth L., Derynck R., Voorhees J. J. Overexpression of transforming growth factor alpha in psoriatic epidermis. Science. 1989 Feb 10;243(4892):811–814. doi: 10.1126/science.2916128. [DOI] [PubMed] [Google Scholar]
  17. Faúndez V., Krauss R., Holuigue L., Garrido J., González A. Epidermal growth factor receptor in synaptic fractions of the rat central nervous system. J Biol Chem. 1992 Oct 5;267(28):20363–20370. [PubMed] [Google Scholar]
  18. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  19. Fisher D. A., Lakshmanan J. Metabolism and effects of epidermal growth factor and related growth factors in mammals. Endocr Rev. 1990 Aug;11(3):418–442. doi: 10.1210/edrv-11-3-418. [DOI] [PubMed] [Google Scholar]
  20. Fukuyama R., Shimizu N. Expression of epidermal growth factor (EGF) and the EGF receptor in human tissues. J Exp Zool. 1991 Jun;258(3):336–343. doi: 10.1002/jez.1402580309. [DOI] [PubMed] [Google Scholar]
  21. Humes H. D., Cieslinski D. A., Coimbra T. M., Messana J. M., Galvao C. Epidermal growth factor enhances renal tubule cell regeneration and repair and accelerates the recovery of renal function in postischemic acute renal failure. J Clin Invest. 1989 Dec;84(6):1757–1761. doi: 10.1172/JCI114359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kajikawa K., Yasui W., Sumiyoshi H., Yoshida K., Nakayama H., Ayhan A., Yokozaki H., Ito H., Tahara E. Expression of epidermal growth factor in human tissues. Immunohistochemical and biochemical analysis. Virchows Arch A Pathol Anat Histopathol. 1991;418(1):27–32. doi: 10.1007/BF01600241. [DOI] [PubMed] [Google Scholar]
  23. Kobrin M. S., Samsoondar J., Kudlow J. E. Alpha-transforming growth factor secreted by untransformed bovine anterior pituitary cells in culture. II. Identification using a sequence-specific monoclonal antibody. J Biol Chem. 1986 Nov 5;261(31):14414–14419. [PubMed] [Google Scholar]
  24. Kommoss F., Wintzer H. O., Von Kleist S., Kohler M., Walker R., Langton B., Van Tran K., Pfleiderer A., Bauknecht T. In situ distribution of transforming growth factor alpha in normal human tissues and in malignant tumours of the ovary. J Pathol. 1990 Nov;162(3):223–230. doi: 10.1002/path.1711620308. [DOI] [PubMed] [Google Scholar]
  25. Lazar L. M., Blum M. Regional distribution and developmental expression of epidermal growth factor and transforming growth factor-alpha mRNA in mouse brain by a quantitative nuclease protection assay. J Neurosci. 1992 May;12(5):1688–1697. doi: 10.1523/JNEUROSCI.12-05-01688.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lee D. C., Rose T. M., Webb N. R., Todaro G. J. Cloning and sequence analysis of a cDNA for rat transforming growth factor-alpha. Nature. 1985 Feb 7;313(6002):489–491. doi: 10.1038/313489a0. [DOI] [PubMed] [Google Scholar]
  27. Lemoine N. R., Hughes C. M., Gullick W. J., Brown C. L., Wynford-Thomas D. Abnormalities of the EGF receptor system in human thyroid neoplasia. Int J Cancer. 1991 Oct 21;49(4):558–561. doi: 10.1002/ijc.2910490414. [DOI] [PubMed] [Google Scholar]
  28. Madtes D. K., Raines E. W., Sakariassen K. S., Assoian R. K., Sporn M. B., Bell G. I., Ross R. Induction of transforming growth factor-alpha in activated human alveolar macrophages. Cell. 1988 Apr 22;53(2):285–293. doi: 10.1016/0092-8674(88)90390-x. [DOI] [PubMed] [Google Scholar]
  29. Massagué J. Transforming growth factor-alpha. A model for membrane-anchored growth factors. J Biol Chem. 1990 Dec 15;265(35):21393–21396. [PubMed] [Google Scholar]
  30. Pascall J. C., Brown K. D. Structural analysis of the 5'-flanking sequence of the mouse epidermal growth factor gene. J Mol Endocrinol. 1988 Jul;1(1):5–11. doi: 10.1677/jme.0.0010005. [DOI] [PubMed] [Google Scholar]
  31. Pascall J. C., Jones D. S., Doel S. M., Clements J. M., Hunter M., Fallon T., Edwards M., Brown K. D. Cloning and characterization of a gene encoding pig epidermal growth factor. J Mol Endocrinol. 1991 Feb;6(1):63–70. doi: 10.1677/jme.0.0060063. [DOI] [PubMed] [Google Scholar]
  32. Pizurki L., Rizzoli R., Caverzasio J., Bonjour J. P. Effect of transforming growth factor-alpha and parathyroid hormone-related protein on phosphate transport in renal cells. Am J Physiol. 1990 Dec;259(6 Pt 2):F929–F935. doi: 10.1152/ajprenal.1990.259.6.F929. [DOI] [PubMed] [Google Scholar]
  33. Rappolee D. A., Brenner C. A., Schultz R., Mark D., Werb Z. Developmental expression of PDGF, TGF-alpha, and TGF-beta genes in preimplantation mouse embryos. Science. 1988 Sep 30;241(4874):1823–1825. doi: 10.1126/science.3175624. [DOI] [PubMed] [Google Scholar]
  34. Rappolee D. A., Mark D., Banda M. J., Werb Z. Wound macrophages express TGF-alpha and other growth factors in vivo: analysis by mRNA phenotyping. Science. 1988 Aug 5;241(4866):708–712. doi: 10.1126/science.3041594. [DOI] [PubMed] [Google Scholar]
  35. Reynolds B. A., Weiss S. Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science. 1992 Mar 27;255(5052):1707–1710. doi: 10.1126/science.1553558. [DOI] [PubMed] [Google Scholar]
  36. Samsoondar J., Kobrin M. S., Kudlow J. E. Alpha-transforming growth factor secreted by untransformed bovine anterior pituitary cells in culture. I. Purification from conditioned medium. J Biol Chem. 1986 Nov 5;261(31):14408–14413. [PubMed] [Google Scholar]
  37. Saunders S. E., Burke J. F. Rapid isolation of miniprep DNA for double strand sequencing. Nucleic Acids Res. 1990 Aug 25;18(16):4948–4948. doi: 10.1093/nar/18.16.4948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schlegel U., Moots P. L., Rosenblum M. K., Thaler H. T., Furneaux H. M. Expression of transforming growth factor alpha in human gliomas. Oncogene. 1990 Dec;5(12):1839–1842. [PubMed] [Google Scholar]
  39. Schultz G., Rotatori D. S., Clark W. EGF and TGF-alpha in wound healing and repair. J Cell Biochem. 1991 Apr;45(4):346–352. doi: 10.1002/jcb.240450407. [DOI] [PubMed] [Google Scholar]
  40. Taub M., Wang Y., Szczesny T. M., Kleinman H. K. Epidermal growth factor or transforming growth factor alpha is required for kidney tubulogenesis in matrigel cultures in serum-free medium. Proc Natl Acad Sci U S A. 1990 May;87(10):4002–4006. doi: 10.1073/pnas.87.10.4002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tokunaga K., Taniguchi H., Yoda K., Shimizu M., Sakiyama S. Nucleotide sequence of a full-length cDNA for mouse cytoskeletal beta-actin mRNA. Nucleic Acids Res. 1986 Mar 25;14(6):2829–2829. doi: 10.1093/nar/14.6.2829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Vassar R., Fuchs E. Transgenic mice provide new insights into the role of TGF-alpha during epidermal development and differentiation. Genes Dev. 1991 May;5(5):714–727. doi: 10.1101/gad.5.5.714. [DOI] [PubMed] [Google Scholar]
  43. Vaughan T. J., James P. S., Pascall J. C., Brown K. D. Expression of the genes for TGF alpha, EGF and the EGF receptor during early pig development. Development. 1992 Nov;116(3):663–669. doi: 10.1242/dev.116.3.663. [DOI] [PubMed] [Google Scholar]
  44. Vaughan T. J., Pascall J. C., Brown K. D. Nucleotide sequence and tissue distribution of mouse transforming growth factor-alpha. Biochim Biophys Acta. 1992 Oct 20;1132(3):322–324. doi: 10.1016/0167-4781(92)90170-5. [DOI] [PubMed] [Google Scholar]
  45. Vaughan T. J., Pascall J. C., James P. S., Brown K. D. Expression of epidermal growth factor and its mRNA in pig kidney, pancreas and other tissues. Biochem J. 1991 Oct 1;279(Pt 1):315–318. doi: 10.1042/bj2790315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Vehaskari V. M., Hering-Smith K. S., Moskowitz D. W., Weiner I. D., Hamm L. L. Effect of epidermal growth factor on sodium transport in the cortical collecting tubule. Am J Physiol. 1989 May;256(5 Pt 2):F803–F809. doi: 10.1152/ajprenal.1989.256.5.F803. [DOI] [PubMed] [Google Scholar]
  47. Werb Z. Expression of EGF and TGF-alpha genes in early mammalian development. Mol Reprod Dev. 1990 Sep;27(1):10–15. doi: 10.1002/mrd.1080270105. [DOI] [PubMed] [Google Scholar]
  48. Wilcox J. N., Derynck R. Localization of cells synthesizing transforming growth factor-alpha mRNA in the mouse brain. J Neurosci. 1988 Jun;8(6):1901–1904. doi: 10.1523/JNEUROSCI.08-06-01901.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wong S. T., Winchell L. F., McCune B. K., Earp H. S., Teixidó J., Massagué J., Herman B., Lee D. C. The TGF-alpha precursor expressed on the cell surface binds to the EGF receptor on adjacent cells, leading to signal transduction. Cell. 1989 Feb 10;56(3):495–506. doi: 10.1016/0092-8674(89)90252-3. [DOI] [PubMed] [Google Scholar]
  50. de Larco J. E., Todaro G. J. Growth factors from murine sarcoma virus-transformed cells. Proc Natl Acad Sci U S A. 1978 Aug;75(8):4001–4005. doi: 10.1073/pnas.75.8.4001. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES