Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1994 Jul 1;13(13):3083–3093. doi: 10.1002/j.1460-2075.1994.tb06607.x

A cell-specific enhancer far upstream of the mouse tyrosinase gene confers high level and copy number-related expression in transgenic mice.

R Ganss 1, L Montoliu 1, A P Monaghan 1, G Schütz 1
PMCID: PMC395199  PMID: 8039502

Abstract

The tyrosinase gene encodes the key enzyme of melanin production and is tightly regulated during development. A yeast artificial chromosome covering the mouse tyrosinase gene has been shown to rescue completely the albino phenotype of recipient mouse strains, conferring copy number-dependent, position-independent expression. To investigate the presence of cis-acting regulatory elements responsible for the appropriate expression of the tyrosinase gene, DNase I hypersensitive site mapping was performed. A melanoma cell-specific DNase I hypersensitive site was identified at -12 kb upstream of the tyrosinase gene. Functional analysis of the corresponding cis-acting element in transgenic mice and transient transfection assays revealed properties of a strong cell-specific enhancer. RNA expression levels of the transgene correlate with copy number, which is reflected in coat colour and eye pigmentation of transgenic mice. Full enhancer activity in transient transfections is obtained with a minimal sequence of 200 bp. Protein binding analysis reveals the presence of a melanoma cell-specific complex which might contribute to the faithful expression of the tyrosinase gene.

Full text

PDF
3083

Images in this article

3084Image
on p.3084
3084Image
on p.3084
3085Image
on p.3085
3086Image
on p.3086
3088Image
on p.3088
3089Image
on p.3089

Selected References

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

  1. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  2. Becker P. B., Ruppert S., Schütz G. Genomic footprinting reveals cell type-specific DNA binding of ubiquitous factors. Cell. 1987 Nov 6;51(3):435–443. doi: 10.1016/0092-8674(87)90639-8. [DOI] [PubMed] [Google Scholar]
  3. Beermann F., Hummler E., Schmid E., Schütz G. Perinatal activation of a tyrosine aminotransferase fusion gene does not occur in albino lethal mice. Mech Dev. 1993 Jul;42(1-2):59–65. doi: 10.1016/0925-4773(93)90098-i. [DOI] [PubMed] [Google Scholar]
  4. Beermann F., Ruppert S., Hummler E., Bosch F. X., Müller G., Rüther U., Schütz G. Rescue of the albino phenotype by introduction of a functional tyrosinase gene into mice. EMBO J. 1990 Sep;9(9):2819–2826. doi: 10.1002/j.1460-2075.1990.tb07470.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beermann F., Schmid E., Schütz G. Expression of the mouse tyrosinase gene during embryonic development: recapitulation of the temporal regulation in transgenic mice. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2809–2813. doi: 10.1073/pnas.89.7.2809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Blom van Assendelft G., Hanscombe O., Grosveld F., Greaves D. R. The beta-globin dominant control region activates homologous and heterologous promoters in a tissue-specific manner. Cell. 1989 Mar 24;56(6):969–977. doi: 10.1016/0092-8674(89)90630-2. [DOI] [PubMed] [Google Scholar]
  7. Bonifer C., Vidal M., Grosveld F., Sippel A. E. Tissue specific and position independent expression of the complete gene domain for chicken lysozyme in transgenic mice. EMBO J. 1990 Sep;9(9):2843–2848. doi: 10.1002/j.1460-2075.1990.tb07473.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Boshart M., Weih F., Schmidt A., Fournier R. E., Schütz G. A cyclic AMP response element mediates repression of tyrosine aminotransferase gene transcription by the tissue-specific extinguisher locus Tse-1. Cell. 1990 Jun 1;61(5):905–916. doi: 10.1016/0092-8674(90)90201-o. [DOI] [PubMed] [Google Scholar]
  9. Bradl M., Klein-Szanto A., Porter S., Mintz B. Malignant melanoma in transgenic mice. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):164–168. doi: 10.1073/pnas.88.1.164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cheyette T. E., Ip T., Faber S., Matsui Y., Chalkley R. Characterization of the factors binding to a PEPCK gene upstream hypersensitive site with LCR activity. Nucleic Acids Res. 1992 Jul 11;20(13):3427–3433. doi: 10.1093/nar/20.13.3427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chung J. H., Whiteley M., Felsenfeld G. A 5' element of the chicken beta-globin domain serves as an insulator in human erythroid cells and protects against position effect in Drosophila. Cell. 1993 Aug 13;74(3):505–514. doi: 10.1016/0092-8674(93)80052-g. [DOI] [PubMed] [Google Scholar]
  12. Crossley M., Orkin S. H. Regulation of the beta-globin locus. Curr Opin Genet Dev. 1993 Apr;3(2):232–237. doi: 10.1016/0959-437x(93)90028-n. [DOI] [PubMed] [Google Scholar]
  13. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Eissenberg J. C., Elgin S. C. Boundary functions in the control of gene expression. Trends Genet. 1991 Oct;7(10):335–340. doi: 10.1016/0168-9525(91)90424-o. [DOI] [PubMed] [Google Scholar]
  15. Elgin S. C. Chromatin structure and gene activity. Curr Opin Cell Biol. 1990 Jun;2(3):437–445. doi: 10.1016/0955-0674(90)90125-x. [DOI] [PubMed] [Google Scholar]
  16. Epner E., Kim C. G., Groudine M. What does the locus control region control? Curr Biol. 1992 May;2(5):262–264. doi: 10.1016/0960-9822(92)90379-o. [DOI] [PubMed] [Google Scholar]
  17. Forrester W. C., Epner E., Driscoll M. C., Enver T., Brice M., Papayannopoulou T., Groudine M. A deletion of the human beta-globin locus activation region causes a major alteration in chromatin structure and replication across the entire beta-globin locus. Genes Dev. 1990 Oct;4(10):1637–1649. doi: 10.1101/gad.4.10.1637. [DOI] [PubMed] [Google Scholar]
  18. Frame M. C., Wilkie N. M., Darling A. J., Chudleigh A., Pintzas A., Lang J. C., Gillespie D. A. Regulation of AP-1/DNA complex formation in vitro. Oncogene. 1991 Feb;6(2):205–209. [PubMed] [Google Scholar]
  19. Gasser S. M., Laemmli U. K. Cohabitation of scaffold binding regions with upstream/enhancer elements of three developmentally regulated genes of D. melanogaster. Cell. 1986 Aug 15;46(4):521–530. doi: 10.1016/0092-8674(86)90877-9. [DOI] [PubMed] [Google Scholar]
  20. Gluecksohn-Waelsch S. Genetic control of morphogenetic and biochemical differentiation: lethal albino deletions in the mouse. Cell. 1979 Feb;16(2):225–237. doi: 10.1016/0092-8674(79)90001-1. [DOI] [PubMed] [Google Scholar]
  21. Greaves D. R., Wilson F. D., Lang G., Kioussis D. Human CD2 3'-flanking sequences confer high-level, T cell-specific, position-independent gene expression in transgenic mice. Cell. 1989 Mar 24;56(6):979–986. doi: 10.1016/0092-8674(89)90631-4. [DOI] [PubMed] [Google Scholar]
  22. Grosveld F., van Assendelft G. B., Greaves D. R., Kollias G. Position-independent, high-level expression of the human beta-globin gene in transgenic mice. Cell. 1987 Dec 24;51(6):975–985. doi: 10.1016/0092-8674(87)90584-8. [DOI] [PubMed] [Google Scholar]
  23. Halaban R., Moellmann G., Tamura A., Kwon B. S., Kuklinska E., Pomerantz S. H., Lerner A. B. Tyrosinases of murine melanocytes with mutations at the albino locus. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7241–7245. doi: 10.1073/pnas.85.19.7241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hardison R., Xu J., Jackson J., Mansberger J., Selifonova O., Grotch B., Biesecker J., Petrykowska H., Miller W. Comparative analysis of the locus control region of the rabbit beta-like gene cluster: HS3 increases transient expression of an embryonic epsilon-globin gene. Nucleic Acids Res. 1993 Mar 11;21(5):1265–1272. doi: 10.1093/nar/21.5.1265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jackson I. J., Bennett D. C. Identification of the albino mutation of mouse tyrosinase by analysis of an in vitro revertant. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7010–7014. doi: 10.1073/pnas.87.18.7010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kellum R., Schedl P. A group of scs elements function as domain boundaries in an enhancer-blocking assay. Mol Cell Biol. 1992 May;12(5):2424–2431. doi: 10.1128/mcb.12.5.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kellum R., Schedl P. A position-effect assay for boundaries of higher order chromosomal domains. Cell. 1991 Mar 8;64(5):941–950. doi: 10.1016/0092-8674(91)90318-s. [DOI] [PubMed] [Google Scholar]
  28. Klüppel M., Beermann F., Ruppert S., Schmid E., Hummler E., Schütz G. The mouse tyrosinase promoter is sufficient for expression in melanocytes and in the pigmented epithelium of the retina. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3777–3781. doi: 10.1073/pnas.88.9.3777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Laemmli U. K., Käs E., Poljak L., Adachi Y. Scaffold-associated regions: cis-acting determinants of chromatin structural loops and functional domains. Curr Opin Genet Dev. 1992 Apr;2(2):275–285. doi: 10.1016/s0959-437x(05)80285-0. [DOI] [PubMed] [Google Scholar]
  30. Lee W., Mitchell P., Tjian R. Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. Cell. 1987 Jun 19;49(6):741–752. doi: 10.1016/0092-8674(87)90612-x. [DOI] [PubMed] [Google Scholar]
  31. Loc P. V., Strätling W. H. The matrix attachment regions of the chicken lysozyme gene co-map with the boundaries of the chromatin domain. EMBO J. 1988 Mar;7(3):655–664. doi: 10.1002/j.1460-2075.1988.tb02860.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Luckow B., Schütz G. CAT constructions with multiple unique restriction sites for the functional analysis of eukaryotic promoters and regulatory elements. Nucleic Acids Res. 1987 Jul 10;15(13):5490–5490. doi: 10.1093/nar/15.13.5490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Montminy M. R., Sevarino K. A., Wagner J. A., Mandel G., Goodman R. H. Identification of a cyclic-AMP-responsive element within the rat somatostatin gene. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6682–6686. doi: 10.1073/pnas.83.18.6682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Müller G., Ruppert S., Schmid E., Schütz G. Functional analysis of alternatively spliced tyrosinase gene transcripts. EMBO J. 1988 Sep;7(9):2723–2730. doi: 10.1002/j.1460-2075.1988.tb03126.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nichols M., Weih F., Schmid W., DeVack C., Kowenz-Leutz E., Luckow B., Boshart M., Schütz G. Phosphorylation of CREB affects its binding to high and low affinity sites: implications for cAMP induced gene transcription. EMBO J. 1992 Sep;11(9):3337–3346. doi: 10.1002/j.1460-2075.1992.tb05412.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Overbeek P. A., Aguilar-Cordova E., Hanten G., Schaffner D. L., Patel P., Lebovitz R. M., Lieberman M. W. Coinjection strategy for visual identification of transgenic mice. Transgenic Res. 1991 Dec;1(1):31–37. doi: 10.1007/BF02512994. [DOI] [PubMed] [Google Scholar]
  38. Palmiter R. D., Brinster R. L. Germ-line transformation of mice. Annu Rev Genet. 1986;20:465–499. doi: 10.1146/annurev.ge.20.120186.002341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Porter S., Larue L., Mintz B. Mosaicism of tyrosinase-locus transcription and chromatin structure in dark vs. light melanocyte clones of homozygous chinchilla-mottled mice. Dev Genet. 1991;12(6):393–402. doi: 10.1002/dvg.1020120604. [DOI] [PubMed] [Google Scholar]
  40. Reitman M., Lee E., Westphal H., Felsenfeld G. Site-independent expression of the chicken beta A-globin gene in transgenic mice. Nature. 1990 Dec 20;348(6303):749–752. doi: 10.1038/348749a0. [DOI] [PubMed] [Google Scholar]
  41. Ruppert S., Müller G., Kwon B., Schütz G. Multiple transcripts of the mouse tyrosinase gene are generated by alternative splicing. EMBO J. 1988 Sep;7(9):2715–2722. doi: 10.1002/j.1460-2075.1988.tb03125.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ryan T. M., Behringer R. R., Martin N. C., Townes T. M., Palmiter R. D., Brinster R. L. A single erythroid-specific DNase I super-hypersensitive site activates high levels of human beta-globin gene expression in transgenic mice. Genes Dev. 1989 Mar;3(3):314–323. doi: 10.1101/gad.3.3.314. [DOI] [PubMed] [Google Scholar]
  43. Schedl A., Beermann F., Thies E., Montoliu L., Kelsey G., Schütz G. Transgenic mice generated by pronuclear injection of a yeast artificial chromosome. Nucleic Acids Res. 1992 Jun 25;20(12):3073–3077. doi: 10.1093/nar/20.12.3073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Schedl A., Larin Z., Montoliu L., Thies E., Kelsey G., Lehrach H., Schütz G. A method for the generation of YAC transgenic mice by pronuclear microinjection. Nucleic Acids Res. 1993 Oct 11;21(20):4783–4787. doi: 10.1093/nar/21.20.4783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Sharpe J. A., Wells D. J., Whitelaw E., Vyas P., Higgs D. R., Wood W. G. Analysis of the human alpha-globin gene cluster in transgenic mice. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11262–11266. doi: 10.1073/pnas.90.23.11262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Steel K. P., Davidson D. R., Jackson I. J. TRP-2/DT, a new early melanoblast marker, shows that steel growth factor (c-kit ligand) is a survival factor. Development. 1992 Aug;115(4):1111–1119. doi: 10.1242/dev.115.4.1111. [DOI] [PubMed] [Google Scholar]
  47. Stewart A. F., Reik A., Schütz G. A simpler and better method to cleave chromatin with DNase 1 for hypersensitive site analyses. Nucleic Acids Res. 1991 Jun 11;19(11):3157–3157. doi: 10.1093/nar/19.11.3157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Stief A., Winter D. M., Strätling W. H., Sippel A. E. A nuclear DNA attachment element mediates elevated and position-independent gene activity. Nature. 1989 Sep 28;341(6240):343–345. doi: 10.1038/341343a0. [DOI] [PubMed] [Google Scholar]
  49. Tabor S., Richardson C. C. Effect of manganese ions on the incorporation of dideoxynucleotides by bacteriophage T7 DNA polymerase and Escherichia coli DNA polymerase I. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4076–4080. doi: 10.1073/pnas.86.11.4076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Talbot D., Grosveld F. The 5'HS2 of the globin locus control region enhances transcription through the interaction of a multimeric complex binding at two functionally distinct NF-E2 binding sites. EMBO J. 1991 Jun;10(6):1391–1398. doi: 10.1002/j.1460-2075.1991.tb07659.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Tanaka S., Yamamoto H., Takeuchi S., Takeuchi T. Melanization in albino mice transformed by introducing cloned mouse tyrosinase gene. Development. 1990 Feb;108(2):223–227. doi: 10.1242/dev.108.2.223. [DOI] [PubMed] [Google Scholar]
  52. Vyas P., Vickers M. A., Simmons D. L., Ayyub H., Craddock C. F., Higgs D. R. Cis-acting sequences regulating expression of the human alpha-globin cluster lie within constitutively open chromatin. Cell. 1992 May 29;69(5):781–793. doi: 10.1016/0092-8674(92)90290-s. [DOI] [PubMed] [Google Scholar]
  53. Wang Y., Macke J. P., Merbs S. L., Zack D. J., Klaunberg B., Bennett J., Gearhart J., Nathans J. A locus control region adjacent to the human red and green visual pigment genes. Neuron. 1992 Sep;9(3):429–440. doi: 10.1016/0896-6273(92)90181-c. [DOI] [PubMed] [Google Scholar]
  54. Wilson C., Bellen H. J., Gehring W. J. Position effects on eukaryotic gene expression. Annu Rev Cell Biol. 1990;6:679–714. doi: 10.1146/annurev.cb.06.110190.003335. [DOI] [PubMed] [Google Scholar]
  55. Yokoyama T., Silversides D. W., Waymire K. G., Kwon B. S., Takeuchi T., Overbeek P. A. Conserved cysteine to serine mutation in tyrosinase is responsible for the classical albino mutation in laboratory mice. Nucleic Acids Res. 1990 Dec 25;18(24):7293–7298. doi: 10.1093/nar/18.24.7293. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES