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. 1993 Jan 1;120(1):253–260. doi: 10.1083/jcb.120.1.253

Extracellular matrix regulates expression of the TGF-beta 1 gene

PMCID: PMC2119480  PMID: 8416992

Abstract

Transforming growth factor-beta (TGF-beta) is a potent regulator of cell proliferation and modulates the interactions of cells with their extracellular matrix (ECM), in part by inducing the synthesis of various ECM proteins. Three different isoforms of TGF-beta are synthesized in a defined pattern in specific cell populations in vivo. In the specific case of TGF-beta 1, this well-defined and limited expression stands in sharp contrast to its synthesis by virtually all cells in culture. Using mammary epithelial cells as a model system, we evaluated the substratum dependence of the expression of TGF-beta 1. The level of TGF-beta 1 expression is high in cells on plastic, but is strongly downregulated when cells are cultured on a reconstituted basement membrane matrix. In contrast, TGF-beta 2 mRNA levels in cells on either substratum remain unchanged. Using the chloramphenicol acetyl transferase gene as reporter gene under the control of the TGF-beta 1 promoter, we show that transcription from this promoter is suppressed when the cells are in contact with either endogenously synthesized or exogenously administered basement membrane. TGF-beta 1 promoter activity is strongly induced by the absence of basement membrane, i.e., by direct contact of the cells with plastic. This modulation of transcription from the TGF-beta 1 promoter occurs in the absence of lactogenic hormones which allow full differentiation. Our results thus indicate that basement membrane is an important regulator of TGF-beta 1 synthesis, and explain why most cells in culture on plastic express TGF- beta 1 in contrast with the more restricted TGF-beta 1 synthesis in vivo. We propose that there is a feedback loop whereby TGF-beta 1- induced synthesis of basement membrane components is repressed once a functional basement membrane is present. Finally, these results together with our current knowledge of regulation of TGF-beta 1 and TGF- beta 2 synthesis, suggest that, in vivo, TGF-beta 1 may play a major role in regulating the ECM synthesis and the cell-ECM interactions, whereas TGF-beta 2 may be more important in morphogenetic processes.

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Selected References

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  1. Aggeler J., Ward J., Blackie L. M., Barcellos-Hoff M. H., Streuli C. H., Bissell M. J. Cytodifferentiation of mouse mammary epithelial cells cultured on a reconstituted basement membrane reveals striking similarities to development in vivo. J Cell Sci. 1991 Jun;99(Pt 2):407–417. doi: 10.1242/jcs.99.2.407. [DOI] [PubMed] [Google Scholar]
  2. Arrick B. A., Korc M., Derynck R. Differential regulation of expression of three transforming growth factor beta species in human breast cancer cell lines by estradiol. Cancer Res. 1990 Jan 15;50(2):299–303. [PubMed] [Google Scholar]
  3. Arrick B. A., Lopez A. R., Elfman F., Ebner R., Damsky C. H., Derynck R. Altered metabolic and adhesive properties and increased tumorigenesis associated with increased expression of transforming growth factor beta 1. J Cell Biol. 1992 Aug;118(3):715–726. doi: 10.1083/jcb.118.3.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barcellos-Hoff M. H., Aggeler J., Ram T. G., Bissell M. J. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane. Development. 1989 Feb;105(2):223–235. doi: 10.1242/dev.105.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bascom C. C., Wolfshohl J. R., Coffey R. J., Jr, Madisen L., Webb N. R., Purchio A. R., Derynck R., Moses H. L. Complex regulation of transforming growth factor beta 1, beta 2, and beta 3 mRNA expression in mouse fibroblasts and keratinocytes by transforming growth factors beta 1 and beta 2. Mol Cell Biol. 1989 Dec;9(12):5508–5515. doi: 10.1128/mcb.9.12.5508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen L. H., Bissell M. J. A novel regulatory mechanism for whey acidic protein gene expression. Cell Regul. 1989 Nov;1(1):45–54. doi: 10.1091/mbc.1.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Derynck R., Jarrett J. A., Chen E. Y., Goeddel D. V. The murine transforming growth factor-beta precursor. J Biol Chem. 1986 Apr 5;261(10):4377–4379. [PubMed] [Google Scholar]
  8. Derynck R., Lindquist P. B., Lee A., Wen D., Tamm J., Graycar J. L., Rhee L., Mason A. J., Miller D. A., Coffey R. J. A new type of transforming growth factor-beta, TGF-beta 3. EMBO J. 1988 Dec 1;7(12):3737–3743. doi: 10.1002/j.1460-2075.1988.tb03257.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Derynck R., Rhee L., Chen E. Y., Van Tilburg A. Intron-exon structure of the human transforming growth factor-beta precursor gene. Nucleic Acids Res. 1987 Apr 10;15(7):3188–3189. doi: 10.1093/nar/15.7.3188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fitzpatrick D. R., Denhez F., Kondaiah P., Akhurst R. J. Differential expression of TGF beta isoforms in murine palatogenesis. Development. 1990 Jul;109(3):585–595. doi: 10.1242/dev.109.3.585. [DOI] [PubMed] [Google Scholar]
  11. Gatherer D., Ten Dijke P., Baird D. T., Akhurst R. J. Expression of TGF-beta isoforms during first trimester human embryogenesis. Development. 1990 Oct;110(2):445–460. doi: 10.1242/dev.110.2.445. [DOI] [PubMed] [Google Scholar]
  12. Geiser A. G., Kim S. J., Roberts A. B., Sporn M. B. Characterization of the mouse transforming growth factor-beta 1 promoter and activation by the Ha-ras oncogene. Mol Cell Biol. 1991 Jan;11(1):84–92. doi: 10.1128/mcb.11.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Glick A. B., Flanders K. C., Danielpour D., Yuspa S. H., Sporn M. B. Retinoic acid induces transforming growth factor-beta 2 in cultured keratinocytes and mouse epidermis. Cell Regul. 1989 Nov;1(1):87–97. doi: 10.1091/mbc.1.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Heine U. I., Munoz E. F., Flanders K. C., Roberts A. B., Sporn M. B. Colocalization of TGF-beta 1 and collagen I and III, fibronectin and glycosaminoglycans during lung branching morphogenesis. Development. 1990 May;109(1):29–36. doi: 10.1242/dev.109.1.29. [DOI] [PubMed] [Google Scholar]
  15. Heine U., Munoz E. F., Flanders K. C., Ellingsworth L. R., Lam H. Y., Thompson N. L., Roberts A. B., Sporn M. B. Role of transforming growth factor-beta in the development of the mouse embryo. J Cell Biol. 1987 Dec;105(6 Pt 2):2861–2876. doi: 10.1083/jcb.105.6.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jakobovits E. B., Schlokat U., Vannice J. L., Derynck R., Levinson A. D. The human transforming growth factor alpha promoter directs transcription initiation from a single site in the absence of a TATA sequence. Mol Cell Biol. 1988 Dec;8(12):5549–5554. doi: 10.1128/mcb.8.12.5549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kim S. J., Glick A., Sporn M. B., Roberts A. B. Characterization of the promoter region of the human transforming growth factor-beta 1 gene. J Biol Chem. 1989 Jan 5;264(1):402–408. [PubMed] [Google Scholar]
  18. Kleinman H. K., McGarvey M. L., Hassell J. R., Star V. L., Cannon F. B., Laurie G. W., Martin G. R. Basement membrane complexes with biological activity. Biochemistry. 1986 Jan 28;25(2):312–318. doi: 10.1021/bi00350a005. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Kähäri V. M., Larjava H., Uitto J. Differential regulation of extracellular matrix proteoglycan (PG) gene expression. Transforming growth factor-beta 1 up-regulates biglycan (PGI), and versican (large fibroblast PG) but down-regulates decorin (PGII) mRNA levels in human fibroblasts in culture. J Biol Chem. 1991 Jun 5;266(16):10608–10615. [PubMed] [Google Scholar]
  21. Lee E. Y., Lee W. H., Kaetzel C. S., Parry G., Bissell M. J. Interaction of mouse mammary epithelial cells with collagen substrata: regulation of casein gene expression and secretion. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1419–1423. doi: 10.1073/pnas.82.5.1419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lee E. Y., Parry G., Bissell M. J. Modulation of secreted proteins of mouse mammary epithelial cells by the collagenous substrata. J Cell Biol. 1984 Jan;98(1):146–155. doi: 10.1083/jcb.98.1.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Li M. L., Aggeler J., Farson D. A., Hatier C., Hassell J., Bissell M. J. Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells. Proc Natl Acad Sci U S A. 1987 Jan;84(1):136–140. doi: 10.1073/pnas.84.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Malipiero U., Höller M., Werner U., Fontana A. Sequence analysis of the promoter region of the glioblastoma derived T cell suppressor factor/transforming growth factor (TGF)-beta 2 gene reveals striking differences to the TGF-beta 1 and -beta 3 genes. Biochem Biophys Res Commun. 1990 Sep 28;171(3):1145–1151. doi: 10.1016/0006-291x(90)90804-v. [DOI] [PubMed] [Google Scholar]
  25. Meager A. Assays for transforming growth factor beta. J Immunol Methods. 1991 Jul 26;141(1):1–14. doi: 10.1016/0022-1759(91)90204-s. [DOI] [PubMed] [Google Scholar]
  26. Millan F. A., Denhez F., Kondaiah P., Akhurst R. J. Embryonic gene expression patterns of TGF beta 1, beta 2 and beta 3 suggest different developmental functions in vivo. Development. 1991 Jan;111(1):131–143. doi: 10.1242/dev.111.1.131. [DOI] [PubMed] [Google Scholar]
  27. Miller D. A., Lee A., Pelton R. W., Chen E. Y., Moses H. L., Derynck R. Murine transforming growth factor-beta 2 cDNA sequence and expression in adult tissues and embryos. Mol Endocrinol. 1989 Jul;3(7):1108–1114. doi: 10.1210/mend-3-7-1108. [DOI] [PubMed] [Google Scholar]
  28. Pelton R. W., Dickinson M. E., Moses H. L., Hogan B. L. In situ hybridization analysis of TGF beta 3 RNA expression during mouse development: comparative studies with TGF beta 1 and beta 2. Development. 1990 Oct;110(2):609–620. doi: 10.1242/dev.110.2.609. [DOI] [PubMed] [Google Scholar]
  29. Pelton R. W., Hogan B. L., Miller D. A., Moses H. L. Differential expression of genes encoding TGFs beta 1, beta 2, and beta 3 during murine palate formation. Dev Biol. 1990 Oct;141(2):456–460. doi: 10.1016/0012-1606(90)90401-4. [DOI] [PubMed] [Google Scholar]
  30. Pelton R. W., Saxena B., Jones M., Moses H. L., Gold L. I. Immunohistochemical localization of TGF beta 1, TGF beta 2, and TGF beta 3 in the mouse embryo: expression patterns suggest multiple roles during embryonic development. J Cell Biol. 1991 Nov;115(4):1091–1105. doi: 10.1083/jcb.115.4.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Robinson S. D., Silberstein G. B., Roberts A. B., Flanders K. C., Daniel C. W. Regulated expression and growth inhibitory effects of transforming growth factor-beta isoforms in mouse mammary gland development. Development. 1991 Nov;113(3):867–878. doi: 10.1242/dev.113.3.867. [DOI] [PubMed] [Google Scholar]
  32. Schmid P., Cox D., Bilbe G., Maier R., McMaster G. K. Differential expression of TGF beta 1, beta 2 and beta 3 genes during mouse embryogenesis. Development. 1991 Jan;111(1):117–130. doi: 10.1242/dev.111.1.117. [DOI] [PubMed] [Google Scholar]
  33. Schmidhauser C., Bissell M. J., Myers C. A., Casperson G. F. Extracellular matrix and hormones transcriptionally regulate bovine beta-casein 5' sequences in stably transfected mouse mammary cells. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9118–9122. doi: 10.1073/pnas.87.23.9118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schmidhauser C., Casperson G. F., Myers C. A., Sanzo K. T., Bolten S., Bissell M. J. A novel transcriptional enhancer is involved in the prolactin- and extracellular matrix-dependent regulation of beta-casein gene expression. Mol Biol Cell. 1992 Jun;3(6):699–709. doi: 10.1091/mbc.3.6.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sieweke M. H., Thompson N. L., Sporn M. B., Bissell M. J. Mediation of wound-related Rous sarcoma virus tumorigenesis by TGF-beta. Science. 1990 Jun 29;248(4963):1656–1660. doi: 10.1126/science.2163544. [DOI] [PubMed] [Google Scholar]
  36. Streuli C. H., Bailey N., Bissell M. J. Control of mammary epithelial differentiation: basement membrane induces tissue-specific gene expression in the absence of cell-cell interaction and morphological polarity. J Cell Biol. 1991 Dec;115(5):1383–1395. doi: 10.1083/jcb.115.5.1383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Streuli C. H., Bissell M. J. Expression of extracellular matrix components is regulated by substratum. J Cell Biol. 1990 Apr;110(4):1405–1415. doi: 10.1083/jcb.110.4.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Thompson N. L., Flanders K. C., Smith J. M., Ellingsworth L. R., Roberts A. B., Sporn M. B. Expression of transforming growth factor-beta 1 in specific cells and tissues of adult and neonatal mice. J Cell Biol. 1989 Feb;108(2):661–669. doi: 10.1083/jcb.108.2.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Van Obberghen-Schilling E., Roche N. S., Flanders K. C., Sporn M. B., Roberts A. B. Transforming growth factor beta 1 positively regulates its own expression in normal and transformed cells. J Biol Chem. 1988 Jun 5;263(16):7741–7746. [PubMed] [Google Scholar]
  40. Werb Z., Tremble P. M., Behrendtsen O., Crowley E., Damsky C. H. Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression. J Cell Biol. 1989 Aug;109(2):877–889. doi: 10.1083/jcb.109.2.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wilcox J. N., Derynck R. Developmental expression of transforming growth factors alpha and beta in mouse fetus. Mol Cell Biol. 1988 Aug;8(8):3415–3422. doi: 10.1128/mcb.8.8.3415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis: a simple method using two oligonucleotide primers and a single-stranded DNA template. DNA. 1984 Dec;3(6):479–488. doi: 10.1089/dna.1.1984.3.479. [DOI] [PubMed] [Google Scholar]

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