Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Sep 1;134(5):1333–1344. doi: 10.1083/jcb.134.5.1333

A potent far-upstream enhancer in the mouse pro alpha 2(I) collagen gene regulates expression of reporter genes in transgenic mice

PMCID: PMC2120987  PMID: 8794872

Abstract

We have identified three DNase I-hypersensitive sites in chromatin between 15 and 17 kb upstream of the mouse pro alpha 2 (I) collagen gene. These sites were detected in cells that produce type I collagen but not in cells that do not express these genes. A construction containing the sequences from -17 kb to +54 bp of the mouse pro alpha 2 (I) collagen gene, cloned upstream of either the Escherichia coli beta- galactosidase or the firefly luciferase reporter gene, showed strong enhancer activity in transgenic mice when compared with the levels seen previously in animals harboring shorter promoter fragments. Especially high levels of expression of the reporter gene were seen in dermis, fascia, and the fibrous layers of many internal organs. High levels of expression could also be detected in some osteoblastic cells. When various fragments of the 5' flanking sequences were cloned upstream of the 350-bp proximal pro alpha 2(I) collagen promoter linked to the lacZ gene, the cis-acting elements responsible for enhancement were localized in the region between -13.5 and -19.5 kb, the same region that contains the three DNase I-hypersensitive sites. Moreover, the DNA segment from -13.5 to -19.5 kb was also able to drive the cell-specific expression of a 220-bp mouse pro alpha 1(I) collagen promoter, which is silent in transgenic mice. Hence, our data suggest that a far-upstream enhancer element plays a role in regulating high levels of expression of the mouse pro alpha 2(I) collagen gene.

Full Text

The Full Text of this article is available as a PDF (6.0 MB).

Selected References

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

  1. Barsh G. S., Roush C. L., Gelinas R. E. DNA and chromatin structure of the human alpha 1 (I) collagen gene. J Biol Chem. 1984 Dec 10;259(23):14906–14913. [PubMed] [Google Scholar]
  2. Beck K. M., Seekamp A. H., Askew G. R., Mei Z., Farrell C. M., Wang S., Lukens L. N. Association of a change in chromatin structure with a tissue-specific switch in transcription start sites in the alpha 2(I) collagen gene. Nucleic Acids Res. 1991 Sep 25;19(18):4975–4982. doi: 10.1093/nar/19.18.4975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Bellard M., Dretzen G., Giangrande A., Ramain P. Nuclease digestion of transcriptionally active chromatin. Methods Enzymol. 1989;170:317–346. doi: 10.1016/0076-6879(89)70054-9. [DOI] [PubMed] [Google Scholar]
  5. Bendeck M. P., Langille B. L. Rapid accumulation of elastin and collagen in the aortas of sheep in the immediate perinatal period. Circ Res. 1991 Oct;69(4):1165–1169. doi: 10.1161/01.res.69.4.1165. [DOI] [PubMed] [Google Scholar]
  6. Bogdanovic Z., Bedalov A., Krebsbach P. H., Pavlin D., Woody C. O., Clark S. H., Thomas H. F., Rowe D. W., Kream B. E., Lichtler A. C. Upstream regulatory elements necessary for expression of the rat COL1A1 promoter in transgenic mice. J Bone Miner Res. 1994 Feb;9(2):285–292. doi: 10.1002/jbmr.5650090218. [DOI] [PubMed] [Google Scholar]
  7. Breindl M., Harbers K., Jaenisch R. Retrovirus-induced lethal mutation in collagen I gene of mice is associated with an altered chromatin structure. Cell. 1984 Aug;38(1):9–16. doi: 10.1016/0092-8674(84)90521-x. [DOI] [PubMed] [Google Scholar]
  8. Chada K., Magram J., Costantini F. An embryonic pattern of expression of a human fetal globin gene in transgenic mice. Nature. 1986 Feb 20;319(6055):685–689. doi: 10.1038/319685a0. [DOI] [PubMed] [Google Scholar]
  9. Cole W. G. Collagen genes: mutations affecting collagen structure and expression. Prog Nucleic Acid Res Mol Biol. 1994;47:29–80. doi: 10.1016/s0079-6603(08)60249-4. [DOI] [PubMed] [Google Scholar]
  10. D'Souza R. N., Niederreither K., de Crombrugghe B. Osteoblast-specific expression of the alpha 2(I) collagen promoter in transgenic mice: correlation with the distribution of TGF-beta 1. J Bone Miner Res. 1993 Sep;8(9):1127–1136. doi: 10.1002/jbmr.5650080914. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Grosveld F., Antoniou M., Berry M., De Boer E., Dillon N., Ellis J., Fraser P., Hanscombe O., Hurst J., Imam A. The regulation of human globin gene switching. Philos Trans R Soc Lond B Biol Sci. 1993 Feb 27;339(1288):183–191. doi: 10.1098/rstb.1993.0015. [DOI] [PubMed] [Google Scholar]
  13. Kaltschmidt C., Muller M., Brem G., Renkawitz R. DNase I hypersensitive sites far upstream of the rat tryptophan oxygenase gene direct developmentally regulated transcription in livers of transgenic mice. Mech Dev. 1994 Mar;45(3):203–210. doi: 10.1016/0925-4773(94)90008-6. [DOI] [PubMed] [Google Scholar]
  14. Khillan J. S., Schmidt A., Overbeek P. A., de Crombrugghe B., Westphal H. Developmental and tissue-specific expression directed by the alpha 2 type I collagen promoter in transgenic mice. Proc Natl Acad Sci U S A. 1986 Feb;83(3):725–729. doi: 10.1073/pnas.83.3.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Liau G., Szapary D., Setoyama C., de Crombrugghe B. Restriction enzyme digestions identify discrete domains in the chromatin around the promoter of the mouse alpha 2(I) collagen gene. J Biol Chem. 1986 Aug 25;261(24):11362–11368. [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. 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]
  19. 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]
  20. 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]
  21. Pihlajaniemi T., Rehn M. Two new collagen subgroups: membrane-associated collagens and types XV and XVII. Prog Nucleic Acid Res Mol Biol. 1995;50:225–262. doi: 10.1016/s0079-6603(08)60816-8. [DOI] [PubMed] [Google Scholar]
  22. Pogulis R. J., Freytag S. O. Contribution of specific cis-acting elements to activity of the mouse pro-alpha 2(I) collagen enhancer. J Biol Chem. 1993 Feb 5;268(4):2493–2499. [PubMed] [Google Scholar]
  23. Rossert J. A., Chen S. S., Eberspaecher H., Smith C. N., de Crombrugghe B. Identification of a minimal sequence of the mouse pro-alpha 1(I) collagen promoter that confers high-level osteoblast expression in transgenic mice and that binds a protein selectively present in osteoblasts. Proc Natl Acad Sci U S A. 1996 Feb 6;93(3):1027–1031. doi: 10.1073/pnas.93.3.1027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rossert J., Eberspaecher H., de Crombrugghe B. 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. J Cell Biol. 1995 Jun;129(5):1421–1432. doi: 10.1083/jcb.129.5.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rossi P., de Crombrugghe B. Identification of a cell-specific transcriptional enhancer in the first intron of the mouse alpha 2 (type I) collagen gene. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5590–5594. doi: 10.1073/pnas.84.16.5590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sudo H., Kodama H. A., Amagai Y., Yamamoto S., Kasai S. In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol. 1983 Jan;96(1):191–198. doi: 10.1083/jcb.96.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Townes T. M., Lingrel J. B., Chen H. Y., Brinster R. L., Palmiter R. D. Erythroid-specific expression of human beta-globin genes in transgenic mice. EMBO J. 1985 Jul;4(7):1715–1723. doi: 10.1002/j.1460-2075.1985.tb03841.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vuust J., Sobel M. E., Martin G. R. Regulation of type I collagen synthesis. Total pro alpha 1(I) and pro alpha 2(I) mRNAs are maintained in a 2:1 ratio under varying rates of collagen synthesis. Eur J Biochem. 1985 Sep 16;151(3):449–453. doi: 10.1111/j.1432-1033.1985.tb09122.x. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Zhong W., Mirkovitch J., Darnell J. E., Jr Tissue-specific regulation of mouse hepatocyte nuclear factor 4 expression. Mol Cell Biol. 1994 Nov;14(11):7276–7284. doi: 10.1128/mcb.14.11.7276. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES