Skip to main content
PMC home page
Molecular Biology of the Cell logoLink to Molecular Biology of the Cell
. 1992 Jun;3(6):699–709. doi: 10.1091/mbc.3.6.699

A novel transcriptional enhancer is involved in the prolactin- and extracellular matrix-dependent regulation of beta-casein gene expression.

C Schmidhauser 1, G F Casperson 1, C A Myers 1, K T Sanzo 1, S Bolten 1, M J Bissell 1
PMCID: PMC275624  PMID: 1498370

Abstract

Lactogenic hormones and extracellular matrix (ECM) act synergistically to regulate beta-casein expression in culture. We have developed a functional subpopulation of the mouse mammary epithelial cell strain COMMA-1D (designated CID 9), which expresses high level of beta-casein, forms alveolar-like structures when plated onto the EHS tumor-derived matrix, and secretes beta-casein unidirectionally into a lumen. We have further shown that ECM- and prolactin-dependent regulations of beta-casein occur mainly at the transcriptional level and that 5' sequences play an important role in these regulations. To address the question of the nature of the DNA sequence requirements for such regulation, we analyzed the bovine beta-casein gene promoter in these cells. We now have located a 160-bp transcriptional enhancer (BCE1) within the 5' flanking region of the beta-casein gene. Using functional assays, we show that BCE1 contains responsive elements for prolactin- and ECM-dependent regulation. BCE1 placed upstream of a truncated and inactive beta-casein promoter (the shortest extending from -89 to +42 bp with regard to the transcription start site) reconstitutes a promoter even more potent than the intact promoter, which contains BCE1 in its normal context more than 1.5 kb upstream. This small fusion promoter also reconstitutes the normal pattern of regulation, including a requirement for both prolactin and ECM and a synergistic action of prolactin and hydrocortisone. By replacing the milk promoter with a heterologous viral promoter, we show that BCE1 participates in the prolactin- and ECM-mediated regulation.

Full text

PDF
700

Images in this article

703Image
on p.703
704Image
on p.704
704Image
on p.704
705Image
on p.705

Selected References

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

  1. Ball R. K., Friis R. R., Schoenenberger C. A., Doppler W., Groner B. Prolactin regulation of beta-casein gene expression and of a cytosolic 120-kd protein in a cloned mouse mammary epithelial cell line. EMBO J. 1988 Jul;7(7):2089–2095. doi: 10.1002/j.1460-2075.1988.tb03048.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baniahmad A., Steiner C., Köhne A. C., Renkawitz R. Modular structure of a chicken lysozyme silencer: involvement of an unusual thyroid hormone receptor binding site. Cell. 1990 May 4;61(3):505–514. doi: 10.1016/0092-8674(90)90532-j. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Bissell D. M., Arenson D. M., Maher J. J., Roll F. J. Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix in normal rat liver. J Clin Invest. 1987 Mar;79(3):801–812. doi: 10.1172/JCI112887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blum J. L., Zeigler M. E., Wicha M. S. Regulation of rat mammary gene expression by extracellular matrix components. Exp Cell Res. 1987 Dec;173(2):322–340. doi: 10.1016/0014-4827(87)90274-6. [DOI] [PubMed] [Google Scholar]
  6. Boutin J. M., Jolicoeur C., Okamura H., Gagnon J., Edery M., Shirota M., Banville D., Dusanter-Fourt I., Djiane J., Kelly P. A. Cloning and expression of the rat prolactin receptor, a member of the growth hormone/prolactin receptor gene family. Cell. 1988 Apr 8;53(1):69–77. doi: 10.1016/0092-8674(88)90488-6. [DOI] [PubMed] [Google Scholar]
  7. Bradley R. S., Brown A. M. The proto-oncogene int-1 encodes a secreted protein associated with the extracellular matrix. EMBO J. 1990 May;9(5):1569–1575. doi: 10.1002/j.1460-2075.1990.tb08276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Dale T. C., Krnacik M. J., Schmidhauser C., Yang C. L., Bissell M. J., Rosen J. M. High-level expression of the rat whey acidic protein gene is mediated by elements in the promoter and 3' untranslated region. Mol Cell Biol. 1992 Mar;12(3):905–914. doi: 10.1128/mcb.12.3.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Danielson K. G., Oborn C. J., Durban E. M., Butel J. S., Medina D. Epithelial mouse mammary cell line exhibiting normal morphogenesis in vivo and functional differentiation in vitro. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3756–3760. doi: 10.1073/pnas.81.12.3756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Davis J. A., Linzer D. I. Expression of multiple forms of the prolactin receptor in mouse liver. Mol Endocrinol. 1989 Apr;3(4):674–680. doi: 10.1210/mend-3-4-674. [DOI] [PubMed] [Google Scholar]
  12. DiPersio C. M., Jackson D. A., Zaret K. S. The extracellular matrix coordinately modulates liver transcription factors and hepatocyte morphology. Mol Cell Biol. 1991 Sep;11(9):4405–4414. doi: 10.1128/mcb.11.9.4405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Doppler W., Groner B., Ball R. K. Prolactin and glucocorticoid hormones synergistically induce expression of transfected rat beta-casein gene promoter constructs in a mammary epithelial cell line. Proc Natl Acad Sci U S A. 1989 Jan;86(1):104–108. doi: 10.1073/pnas.86.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Doppler W., Höck W., Hofer P., Groner B., Ball R. K. Prolactin and glucocorticoid hormones control transcription of the beta-casein gene by kinetically distinct mechanisms. Mol Endocrinol. 1990 Jun;4(6):912–919. doi: 10.1210/mend-4-6-912. [DOI] [PubMed] [Google Scholar]
  15. Doppler W., Villunger A., Jennewein P., Brduscha K., Groner B., Ball R. K. Lactogenic hormone and cell type-specific control of the whey acidic protein gene promoter in transfected mouse cells. Mol Endocrinol. 1991 Nov;5(11):1624–1632. doi: 10.1210/mend-5-11-1624. [DOI] [PubMed] [Google Scholar]
  16. Edery M., Jolicoeur C., Levi-Meyrueis C., Dusanter-Fourt I., Pétridou B., Boutin J. M., Lesueur L., Kelly P. A., Djiane J. Identification and sequence analysis of a second form of prolactin receptor by molecular cloning of complementary DNA from rabbit mammary gland. Proc Natl Acad Sci U S A. 1989 Mar;86(6):2112–2116. doi: 10.1073/pnas.86.6.2112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Emerman J. T., Enami J., Pitelka D. R., Nandi S. Hormonal effects on intracellular and secreted casein in cultures of mouse mammary epithelial cells on floating collagen membranes. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4466–4470. doi: 10.1073/pnas.74.10.4466. [DOI] [PMC free article] [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. Friedman S. L., Roll F. J., Boyles J., Arenson D. M., Bissell D. M. Maintenance of differentiated phenotype of cultured rat hepatic lipocytes by basement membrane matrix. J Biol Chem. 1989 Jun 25;264(18):10756–10762. [PubMed] [Google Scholar]
  20. Guyette W. A., Matusik R. J., Rosen J. M. Prolactin-mediated transcriptional and post-transcriptional control of casein gene expression. Cell. 1979 Aug;17(4):1013–1023. doi: 10.1016/0092-8674(79)90340-4. [DOI] [PubMed] [Google Scholar]
  21. Highkin M. K., Krivi G. G., Hippenmeyer P. J. Characterization and comparison of avian and murine helper cell lines for production of replication-defective retroviruses for avian transformation. Poult Sci. 1991 Apr;70(4):970–981. doi: 10.3382/ps.0700970. [DOI] [PubMed] [Google Scholar]
  22. Kageyama R., Pastan I. Molecular cloning and characterization of a human DNA binding factor that represses transcription. Cell. 1989 Dec 1;59(5):815–825. doi: 10.1016/0092-8674(89)90605-3. [DOI] [PubMed] [Google Scholar]
  23. Klagsbrun M. The affinity of fibroblast growth factors (FGFs) for heparin; FGF-heparan sulfate interactions in cells and extracellular matrix. Curr Opin Cell Biol. 1990 Oct;2(5):857–863. doi: 10.1016/0955-0674(90)90084-r. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Lesueur L., Edery M., Ali S., Paly J., Kelly P. A., Djiane J. Comparison of long and short forms of the prolactin receptor on prolactin-induced milk protein gene transcription. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):824–828. doi: 10.1073/pnas.88.3.824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Liu J. K., DiPersio C. M., Zaret K. S. Extracellular signals that regulate liver transcription factors during hepatic differentiation in vitro. Mol Cell Biol. 1991 Feb;11(2):773–784. doi: 10.1128/mcb.11.2.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Majors J., Varmus H. E. A small region of the mouse mammary tumor virus long terminal repeat confers glucocorticoid hormone regulation on a linked heterologous gene. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5866–5870. doi: 10.1073/pnas.80.19.5866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rathjen P. D., Toth S., Willis A., Heath J. K., Smith A. G. Differentiation inhibiting activity is produced in matrix-associated and diffusible forms that are generated by alternate promoter usage. Cell. 1990 Sep 21;62(6):1105–1114. doi: 10.1016/0092-8674(90)90387-t. [DOI] [PubMed] [Google Scholar]
  30. Rojkind M., Gatmaitan Z., Mackensen S., Giambrone M. A., Ponce P., Reid L. M. Connective tissue biomatrix: its isolation and utilization for long-term cultures of normal rat hepatocytes. J Cell Biol. 1980 Oct;87(1):255–263. doi: 10.1083/jcb.87.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Saffer J. D., Jackson S. P., Annarella M. B. Developmental expression of Sp1 in the mouse. Mol Cell Biol. 1991 Apr;11(4):2189–2199. doi: 10.1128/mcb.11.4.2189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Schmitt-Ney M., Doppler W., Ball R. K., Groner B. Beta-casein gene promoter activity is regulated by the hormone-mediated relief of transcriptional repression and a mammary-gland-specific nuclear factor. Mol Cell Biol. 1991 Jul;11(7):3745–3755. doi: 10.1128/mcb.11.7.3745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. 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]
  36. Taketani Y., Oka T. Tumor promoter 12-O-tetradecanoylphorbol 13-acetate, like epidermal growth factor, stimulates cell proliferation and inhibits differentiation of mouse mammary epithelial cells in culture. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1646–1649. doi: 10.1073/pnas.80.6.1646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Topper Y. J., Freeman C. S. Multiple hormone interactions in the developmental biology of the mammary gland. Physiol Rev. 1980 Oct;60(4):1049–1106. doi: 10.1152/physrev.1980.60.4.1049. [DOI] [PubMed] [Google Scholar]
  39. Watson C. J., Gordon K. E., Robertson M., Clark A. J. Interaction of DNA-binding proteins with a milk protein gene promoter in vitro: identification of a mammary gland-specific factor. Nucleic Acids Res. 1991 Dec 11;19(23):6603–6610. doi: 10.1093/nar/19.23.6603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wicha M. S., Lowrie G., Kohn E., Bagavandoss P., Mahn T. Extracellular matrix promotes mammary epithelial growth and differentiation in vitro. Proc Natl Acad Sci U S A. 1982 May;79(10):3213–3217. doi: 10.1073/pnas.79.10.3213. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular Biology of the Cell are provided here courtesy of American Society for Cell Biology

RESOURCES