Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1995 Jun 1;129(5):1421–1432. doi: 10.1083/jcb.129.5.1421

Separate cis-acting DNA elements of the mouse pro-alpha 1(I) collagen promoter direct expression of reporter genes to different type I collagen-producing cells in transgenic mice

PMCID: PMC2120462  PMID: 7775585

Abstract

The genes coding for the two type I collagen chains, which are active selectively in osteoblasts, odontoblasts, fibroblasts, and some mesenchymal cells, constitute good models for studying the mechanisms responsible for the cell-specific activity of genes which are expressed in a small number of discrete cell types. To test whether separate genetic elements could direct the activity of the mouse pro-alpha 1(I) collagen gene to different cell types in which it is expressed, transgenic mice were generated harboring various fragments of the proximal promoter of this gene cloned upstream of the Escherichia coli beta-galactosidase gene. During embryonic development, X-gal staining allows for the precise identification of the different cell types in which the beta-galactosidase gene is active. Transgenic mice harboring 900 bp of the pro-alpha 1(I) proximal promoter expressed the transgene at relatively low levels almost exclusively in skin. In mice containing 2.3 kb of this proximal promoter, the transgene was also expressed at high levels in osteoblasts and odontoblasts, but not in other type I collagen-producing cells. Transgenic mice harboring 3.2 kb of the proximal promoter showed an additional high level expression of the transgene in tendon and fascia fibroblasts. The pattern of expression of the lacZ transgene directed by the 0.9- and 2.3-kb pro-alpha 1(I) proximal promoters was confirmed by using the firefly luciferase gene as a reporter gene. The pattern of expression of this transgene, which can be detected even when it is active at very low levels, paralleled that of the beta-galactosidase gene. These data strongly suggest a modular arrangement of separate cell-specific cis-acting elements that can activate the mouse pro-alpha(I) collagen gene in different type I collagen-producing cells. At least three different types of cell- specific elements would be located in the first 3.2 kb of the promoter: (a) an element that confers low level expression in dermal fibroblasts; (b) a second that mediates high level expression in osteoblasts and odontoblasts; and (c) one responsible for high level expression in tendon and fascia fibroblasts. Our data also imply that other cis- acting cell-specific elements which direct activity of the gene to still other type I collagen-producing cells remain to be identified.

Full Text

The Full Text of this article is available as a PDF (8.9 MB).

Selected References

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

  1. Bedalov A., Breault D. T., Sokolov B. P., Lichtler A. C., Bedalov I., Clark S. H., Mack K., Khillan J. S., Woody C. O., Kream B. E. Regulation of the alpha 1(I) collagen promoter in vascular smooth muscle cells. Comparison with other alpha 1(I) collagen-producing cells in transgenic animals and cultured cells. J Biol Chem. 1994 Feb 18;269(7):4903–4909. [PubMed] [Google Scholar]
  2. Bornstein P., Sage H. Regulation of collagen gene expression. Prog Nucleic Acid Res Mol Biol. 1989;37:67–106. doi: 10.1016/s0079-6603(08)60695-9. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Goldberg H., Helaakoski T., Garrett L. A., Karsenty G., Pellegrino A., Lozano G., Maity S., de Crombrugghe B. Tissue-specific expression of the mouse alpha 2(I) collagen promoter. Studies in transgenic mice and in tissue culture cells. J Biol Chem. 1992 Sep 25;267(27):19622–19630. [PubMed] [Google Scholar]
  5. Harbers K., Kuehn M., Delius H., Jaenisch R. Insertion of retrovirus into the first intron of alpha 1(I) collagen gene to embryonic lethal mutation in mice. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1504–1508. doi: 10.1073/pnas.81.5.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hartung S., Jaenisch R., Breindl M. Retrovirus insertion inactivates mouse alpha 1(I) collagen gene by blocking initiation of transcription. 1986 Mar 27-Apr 2Nature. 320(6060):365–367. doi: 10.1038/320365a0. [DOI] [PubMed] [Google Scholar]
  7. Karsenty G., de Crombrugghe B. Two different negative and one positive regulatory factors interact with a short promoter segment of the alpha 1 (I) collagen gene. J Biol Chem. 1990 Jun 15;265(17):9934–9942. [PubMed] [Google Scholar]
  8. Kratochwil K., von der Mark K., Kollar E. J., Jaenisch R., Mooslehner K., Schwarz M., Haase K., Gmachl I., Harbers K. Retrovirus-induced insertional mutation in Mov13 mice affects collagen I expression in a tissue-specific manner. Cell. 1989 Jun 2;57(5):807–816. doi: 10.1016/0092-8674(89)90795-2. [DOI] [PubMed] [Google Scholar]
  9. Liska D. J., Reed M. J., Sage E. H., Bornstein P. Cell-specific expression of alpha 1(I) collagen-hGH minigenes in transgenic mice. J Cell Biol. 1994 May;125(3):695–704. doi: 10.1083/jcb.125.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McKnight R. A., Shamay A., Sankaran L., Wall R. J., Hennighausen L. Matrix-attachment regions can impart position-independent regulation of a tissue-specific gene in transgenic mice. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6943–6947. doi: 10.1073/pnas.89.15.6943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McLeod M. J. Differential staining of cartilage and bone in whole mouse fetuses by alcian blue and alizarin red S. Teratology. 1980 Dec;22(3):299–301. doi: 10.1002/tera.1420220306. [DOI] [PubMed] [Google Scholar]
  12. Mercer E. H., Hoyle G. W., Kapur R. P., Brinster R. L., Palmiter R. D. The dopamine beta-hydroxylase gene promoter directs expression of E. coli lacZ to sympathetic and other neurons in adult transgenic mice. Neuron. 1991 Nov;7(5):703–716. doi: 10.1016/0896-6273(91)90274-4. [DOI] [PubMed] [Google Scholar]
  13. Niederreither K., D'Souza R. N., de Crombrugghe B. Minimal DNA sequences that control the cell lineage-specific expression of the pro alpha 2(I) collagen promoter in transgenic mice. J Cell Biol. 1992 Dec;119(5):1361–1370. doi: 10.1083/jcb.119.5.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pavlin D., Lichtler A. C., Bedalov A., Kream B. E., Harrison J. R., Thomas H. F., Gronowicz G. A., Clark S. H., Woody C. O., Rowe D. W. Differential utilization of regulatory domains within the alpha 1(I) collagen promoter in osseous and fibroblastic cells. J Cell Biol. 1992 Jan;116(1):227–236. doi: 10.1083/jcb.116.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schwarz M., Harbers K., Kratochwil K. Transcription of a mutant collagen I gene is a cell type and stage-specific marker for odontoblast and osteoblast differentiation. Development. 1990 Apr;108(4):717–726. doi: 10.1242/dev.108.4.717. [DOI] [PubMed] [Google Scholar]
  16. Slack J. L., Liska D. J., Bornstein P. An upstream regulatory region mediates high-level, tissue-specific expression of the human alpha 1(I) collagen gene in transgenic mice. Mol Cell Biol. 1991 Apr;11(4):2066–2074. doi: 10.1128/mcb.11.4.2066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sokolov B. P., Mays P. K., Khillan J. S., Prockop D. J. Tissue- and development-specific expression in transgenic mice of a type I procollagen (COL1A1) minigene construct with 2.3 kb of the promoter region and 2 kb of the 3'-flanking region. Specificity is independent of the putative regulatory sequences in the first intron. Biochemistry. 1993 Sep 7;32(35):9242–9249. doi: 10.1021/bi00086a033. [DOI] [PubMed] [Google Scholar]
  18. Wood W. M., Kao M. Y., Gordon D. F., Ridgway E. C. Thyroid hormone regulates the mouse thyrotropin beta-subunit gene promoter in transfected primary thyrotropes. J Biol Chem. 1989 Sep 5;264(25):14840–14847. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES