Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1990 Feb;10(2):760–769. doi: 10.1128/mcb.10.2.760

Highly conserved upstream regions of the alpha 1-antitrypsin gene in two mouse species govern liver-specific expression by different mechanisms.

J J Latimer 1, F G Berger 1, H Baumann 1
PMCID: PMC360876  PMID: 1689000

Abstract

alpha 1-Antitrypsin (AT), the major elastase inhibitor in mammalian serum, is produced primarily in the liver. We have characterized AT gene structure and expression in the mouse species Mus caroli, which expresses high levels of AT in the kidneys as well as in the liver. Analysis of cDNA and genomic clones showed that the AT gene in M. caroli exhibits high sequence homology (greater than 90%) to the gene in laboratory mice (M. domesticus) throughout the coding and 5'-flanking regions. Despite this extensive sequence conservation, the functional organization of cis-acting regulatory elements governing liver-specific expression is strikingly different between these species. Transient-transfection assays showed that the proximal region of the M. caroli promoter (i.e., between -120 and -2 relative to the transcriptional start site) is 10-fold more active than the analogous region of M. domesticus in driving the expression of an indicator gene in cultured liver cells. The increased activity of the proximal region of the M. caroli AT promoter appears to be the result of one or both of the two base substitutions at positions -46 and -48. The weak proximal promoter in M. domesticus is compensated for by the presence of upstream, liver-specific enhancers between -199 and -520; the analogous region in M. caroli is inactive. Thus, during the course of evolution, the modest 7% sequence divergence that has occurred between the 5'-flanking regions of the AT genes in these two species has generated distinct, yet equally effective, modes of hepatocyte-specific expression.

Full text

PDF
762

Images in this article

764Image
on p.764
765Image
on p.765
768Image
on p.768

Selected References

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

  1. Barth R. K., Gross K. W., Gremke L. C., Hastie N. D. Developmentally regulated mRNAs in mouse liver. Proc Natl Acad Sci U S A. 1982 Jan;79(2):500–504. doi: 10.1073/pnas.79.2.500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berger F. G., Baumann H. An evolutionary switch in tissue-specific gene expression. Abundant expression of alpha 1-antitrypsin in the kidney of a wild mouse species. J Biol Chem. 1985 Jan 25;260(2):1160–1165. [PubMed] [Google Scholar]
  3. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  4. Chapman V., Forrester L., Sanford J., Hastie N., Rossant J. Cell lineage-specific undermethylation of mouse repetitive DNA. Nature. 1984 Jan 19;307(5948):284–286. doi: 10.1038/307284a0. [DOI] [PubMed] [Google Scholar]
  5. Ciliberto G., Dente L., Cortese R. Cell-specific expression of a transfected human alpha 1-antitrypsin gene. Cell. 1985 Jun;41(2):531–540. doi: 10.1016/s0092-8674(85)80026-x. [DOI] [PubMed] [Google Scholar]
  6. Costa R. H., Grayson D. R., Darnell J. E., Jr Multiple hepatocyte-enriched nuclear factors function in the regulation of transthyretin and alpha 1-antitrypsin genes. Mol Cell Biol. 1989 Apr;9(4):1415–1425. doi: 10.1128/mcb.9.4.1415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Simone V., Ciliberto G., Hardon E., Paonessa G., Palla F., Lundberg L., Cortese R. Cis- and trans-acting elements responsible for the cell-specific expression of the human alpha 1-antitrypsin gene. EMBO J. 1987 Sep;6(9):2759–2766. doi: 10.1002/j.1460-2075.1987.tb02570.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Field L. J., Philbrick W. M., Howles P. N., Dickinson D. P., McGowan R. A., Gross K. W. Expression of tissue-specific Ren-1 and Ren-2 genes of mice: comparative analysis of 5'-proximal flanking regions. Mol Cell Biol. 1984 Nov;4(11):2321–2331. doi: 10.1128/mcb.4.11.2321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  11. Grayson D. R., Costa R. H., Xanthopoulos K. G., Darnell J. E., Jr A cell-specific enhancer of the mouse alpha 1-antitrypsin gene has multiple functional regions and corresponding protein-binding sites. Mol Cell Biol. 1988 Mar;8(3):1055–1066. doi: 10.1128/mcb.8.3.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  13. Hall C. V., Jacob P. E., Ringold G. M., Lee F. Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells. J Mol Appl Genet. 1983;2(1):101–109. [PubMed] [Google Scholar]
  14. Held W. A., West K., Gallagher J. F. Importance of initiation factor preparations in the translation of reovirus and globin mRNAs lacking a 5'-terminal 7-methylguanosine. J Biol Chem. 1977 Dec 10;252(23):8489–8497. [PubMed] [Google Scholar]
  15. Hill R. E., Shaw P. H., Boyd P. A., Baumann H., Hastie N. D. Plasma protease inhibitors in mouse and man: divergence within the reactive centre regions. Nature. 1984 Sep 13;311(5982):175–177. doi: 10.1038/311175a0. [DOI] [PubMed] [Google Scholar]
  16. Krauter K. S., Citron B. A., Hsu M. T., Powell D., Darnell J. E., Jr Isolation and characterization of the alpha 1-antitrypsin gene of mice. DNA. 1986 Feb;5(1):29–36. doi: 10.1089/dna.1986.5.29. [DOI] [PubMed] [Google Scholar]
  17. Labarca C., Paigen K. MRNA-directed synthesis of catalytically active mouse beta-glucuronidase in Xenopus oocytes. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4462–4465. doi: 10.1073/pnas.74.10.4462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Latimer J. J., Berger F. G., Baumann H. Developmental expression, cellular localization, and testosterone regulation of alpha 1-antitrypsin in Mus caroli kidney. J Biol Chem. 1987 Sep 15;262(26):12641–12646. [PMC free article] [PubMed] [Google Scholar]
  19. Long G. L., Chandra T., Woo S. L., Davie E. W., Kurachi K. Complete sequence of the cDNA for human alpha 1-antitrypsin and the gene for the S variant. Biochemistry. 1984 Oct 9;23(21):4828–4837. doi: 10.1021/bi00316a003. [DOI] [PubMed] [Google Scholar]
  20. Maniatis T., Goodbourn S., Fischer J. A. Regulation of inducible and tissue-specific gene expression. Science. 1987 Jun 5;236(4806):1237–1245. doi: 10.1126/science.3296191. [DOI] [PubMed] [Google Scholar]
  21. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  22. Perlino E., Cortese R., Ciliberto G. The human alpha 1-antitrypsin gene is transcribed from two different promoters in macrophages and hepatocytes. EMBO J. 1987 Sep;6(9):2767–2771. doi: 10.1002/j.1460-2075.1987.tb02571.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rheaume C., Latimer J. J., Baumann H., Berger F. G. Tissue- and species-specific regulation of murine alpha 1-antitrypsin gene transcription. J Biol Chem. 1988 Oct 15;263(29):15118–15121. [PMC free article] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shen R. F., Li Y., Sifers R. N., Wang H., Hardick C., Tsai S. Y., Woo S. L. Tissue-specific expression of the human alpha 1-antitrypsin gene is controlled by multiple cis-regulatory elements. Nucleic Acids Res. 1987 Oct 26;15(20):8399–8415. doi: 10.1093/nar/15.20.8399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sifers R. N., Carlson J. A., Clift S. M., DeMayo F. J., Bullock D. W., Woo S. L. Tissue specific expression of the human alpha-1-antitrypsin gene in transgenic mice. Nucleic Acids Res. 1987 Feb 25;15(4):1459–1475. doi: 10.1093/nar/15.4.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  28. Travis J., Salvesen G. S. Human plasma proteinase inhibitors. Annu Rev Biochem. 1983;52:655–709. doi: 10.1146/annurev.bi.52.070183.003255. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES