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. 1997 Jul;17(7):3497–3507. doi: 10.1128/mcb.17.7.3497

Transactivation domains facilitate promoter occupancy for the dioxin-inducible CYP1A1 gene in vivo.

H P Ko 1, S T Okino 1, Q Ma 1, J P Whitlock Jr 1
PMCID: PMC232203  PMID: 9199285

Abstract

We have studied the transcriptional regulation of the dioxin-inducible mouse CYP1A1 gene in its native chromosomal setting. We analyzed the ability of aromatic hydrocarbon receptor (AhR) mutants and AhR chimeras to restore dioxin responsiveness to the CYP1A1 gene in AhR-defective mouse hepatoma cells. Our data reveal that transactivation domains in AhR's C-terminal half mediate occupancy of the nuclear factor 1 site and TATA box for the CYP1A1 promoter in vivo. Transactivation domains of VP16 and AhR nuclear translocator, but not Sp1, can substitute for AhR's C-terminal half in facilitating protein binding at the promoter. Our data also reveal an apparent linear relationship between promoter occupancy and CYP1A1 gene expression in chromatin. These findings provide new insights into the in vivo mechanism of transcriptional activation for an interesting mammalian gene.

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Selected References

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  1. Bannister A. J., Kouzarides T. The CBP co-activator is a histone acetyltransferase. Nature. 1996 Dec 19;384(6610):641–643. doi: 10.1038/384641a0. [DOI] [PubMed] [Google Scholar]
  2. Blau J., Xiao H., McCracken S., O'Hare P., Greenblatt J., Bentley D. Three functional classes of transcriptional activation domain. Mol Cell Biol. 1996 May;16(5):2044–2055. doi: 10.1128/mcb.16.5.2044. [DOI] [PMC free article] [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. Burbach K. M., Poland A., Bradfield C. A. Cloning of the Ah-receptor cDNA reveals a distinctive ligand-activated transcription factor. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8185–8189. doi: 10.1073/pnas.89.17.8185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Byrne G. W., Ruddle F. H. Multiplex gene regulation: a two-tiered approach to transgene regulation in transgenic mice. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5473–5477. doi: 10.1073/pnas.86.14.5473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cairns B. R., Lorch Y., Li Y., Zhang M., Lacomis L., Erdjument-Bromage H., Tempst P., Du J., Laurent B., Kornberg R. D. RSC, an essential, abundant chromatin-remodeling complex. Cell. 1996 Dec 27;87(7):1249–1260. doi: 10.1016/s0092-8674(00)81820-6. [DOI] [PubMed] [Google Scholar]
  7. Chatterjee S., Struhl K. Connecting a promoter-bound protein to TBP bypasses the need for a transcriptional activation domain. Nature. 1995 Apr 27;374(6525):820–822. doi: 10.1038/374820a0. [DOI] [PubMed] [Google Scholar]
  8. Cress W. D., Triezenberg S. J. Critical structural elements of the VP16 transcriptional activation domain. Science. 1991 Jan 4;251(4989):87–90. doi: 10.1126/science.1846049. [DOI] [PubMed] [Google Scholar]
  9. Dang C. V., Barrett J., Villa-Garcia M., Resar L. M., Kato G. J., Fearon E. R. Intracellular leucine zipper interactions suggest c-Myc hetero-oligomerization. Mol Cell Biol. 1991 Feb;11(2):954–962. doi: 10.1128/mcb.11.2.954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Denison M. S., Whitlock J. P., Jr Xenobiotic-inducible transcription of cytochrome P450 genes. J Biol Chem. 1995 Aug 4;270(31):18175–18178. doi: 10.1074/jbc.270.31.18175. [DOI] [PubMed] [Google Scholar]
  11. Dranoff G., Jaffee E., Lazenby A., Golumbek P., Levitsky H., Brose K., Jackson V., Hamada H., Pardoll D., Mulligan R. C. Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent, specific, and long-lasting anti-tumor immunity. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3539–3543. doi: 10.1073/pnas.90.8.3539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Durrin L. K., Whitlock J. P., Jr 2,3,7,8-Tetrachlorodibenzo-p-dioxin-inducible aryl hydrocarbon receptor-mediated change in CYP1A1 chromatin structure occurs independently of transcription. Mol Cell Biol. 1989 Dec;9(12):5733–5737. doi: 10.1128/mcb.9.12.5733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Emami K. H., Navarre W. W., Smale S. T. Core promoter specificities of the Sp1 and VP16 transcriptional activation domains. Mol Cell Biol. 1995 Nov;15(11):5906–5916. doi: 10.1128/mcb.15.11.5906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fearon E. R., Finkel T., Gillison M. L., Kennedy S. P., Casella J. F., Tomaselli G. F., Morrow J. S., Van Dang C. Karyoplasmic interaction selection strategy: a general strategy to detect protein-protein interactions in mammalian cells. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7958–7962. doi: 10.1073/pnas.89.17.7958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fisher J. M., Wu L., Denison M. S., Whitlock J. P., Jr Organization and function of a dioxin-responsive enhancer. J Biol Chem. 1990 Jun 15;265(17):9676–9681. [PubMed] [Google Scholar]
  16. Gerber H. P., Hagmann M., Seipel K., Georgiev O., West M. A., Litingtung Y., Schaffner W., Corden J. L. RNA polymerase II C-terminal domain required for enhancer-driven transcription. Nature. 1995 Apr 13;374(6523):660–662. doi: 10.1038/374660a0. [DOI] [PubMed] [Google Scholar]
  17. Goodrich J. A., Cutler G., Tjian R. Contacts in context: promoter specificity and macromolecular interactions in transcription. Cell. 1996 Mar 22;84(6):825–830. doi: 10.1016/s0092-8674(00)81061-2. [DOI] [PubMed] [Google Scholar]
  18. Goodrich J. A., Hoey T., Thut C. J., Admon A., Tjian R. Drosophila TAFII40 interacts with both a VP16 activation domain and the basal transcription factor TFIIB. Cell. 1993 Nov 5;75(3):519–530. doi: 10.1016/0092-8674(93)90386-5. [DOI] [PubMed] [Google Scholar]
  19. Hahn S. Structure(?) and function of acidic transcription activators. Cell. 1993 Feb 26;72(4):481–483. doi: 10.1016/0092-8674(93)90064-w. [DOI] [PubMed] [Google Scholar]
  20. Hankinson O. Single-step selection of clones of a mouse hepatoma line deficient in aryl hydrocarbon hydroxylase. Proc Natl Acad Sci U S A. 1979 Jan;76(1):373–376. doi: 10.1073/pnas.76.1.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hankinson O. The aryl hydrocarbon receptor complex. Annu Rev Pharmacol Toxicol. 1995;35:307–340. doi: 10.1146/annurev.pa.35.040195.001515. [DOI] [PubMed] [Google Scholar]
  22. Hoffman E. C., Reyes H., Chu F. F., Sander F., Conley L. H., Brooks B. A., Hankinson O. Cloning of a factor required for activity of the Ah (dioxin) receptor. Science. 1991 May 17;252(5008):954–958. doi: 10.1126/science.1852076. [DOI] [PubMed] [Google Scholar]
  23. Horikoshi M., Hai T., Lin Y. S., Green M. R., Roeder R. G. Transcription factor ATF interacts with the TATA factor to facilitate establishment of a preinitiation complex. Cell. 1988 Sep 23;54(7):1033–1042. doi: 10.1016/0092-8674(88)90118-3. [DOI] [PubMed] [Google Scholar]
  24. Huang Z. J., Edery I., Rosbash M. PAS is a dimerization domain common to Drosophila period and several transcription factors. Nature. 1993 Jul 15;364(6434):259–262. doi: 10.1038/364259a0. [DOI] [PubMed] [Google Scholar]
  25. Ingles C. J., Shales M., Cress W. D., Triezenberg S. J., Greenblatt J. Reduced binding of TFIID to transcriptionally compromised mutants of VP16. Nature. 1991 Jun 13;351(6327):588–590. doi: 10.1038/351588a0. [DOI] [PubMed] [Google Scholar]
  26. Israel D. I., Whitlock J. P., Jr Induction of mRNA specific for cytochrome P1-450 in wild type and variant mouse hepatoma cells. J Biol Chem. 1983 Sep 10;258(17):10390–10394. [PubMed] [Google Scholar]
  27. Jain S., Dolwick K. M., Schmidt J. V., Bradfield C. A. Potent transactivation domains of the Ah receptor and the Ah receptor nuclear translocator map to their carboxyl termini. J Biol Chem. 1994 Dec 16;269(50):31518–31524. [PubMed] [Google Scholar]
  28. Jones K. W., Whitlock J. P., Jr Functional analysis of the transcriptional promoter for the CYP1A1 gene. Mol Cell Biol. 1990 Oct;10(10):5098–5105. doi: 10.1128/mcb.10.10.5098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kadonaga J. T., Carner K. R., Masiarz F. R., Tjian R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell. 1987 Dec 24;51(6):1079–1090. doi: 10.1016/0092-8674(87)90594-0. [DOI] [PubMed] [Google Scholar]
  30. Klages N., Strubin M. Stimulation of RNA polymerase II transcription initiation by recruitment of TBP in vivo. Nature. 1995 Apr 27;374(6525):822–823. doi: 10.1038/374822a0. [DOI] [PubMed] [Google Scholar]
  31. Klein C., Struhl K. Increased recruitment of TATA-binding protein to the promoter by transcriptional activation domains in vivo. Science. 1994 Oct 14;266(5183):280–282. doi: 10.1126/science.7939664. [DOI] [PubMed] [Google Scholar]
  32. Ko H. P., Okino S. T., Ma Q., Whitlock J. P., Jr Dioxin-induced CYP1A1 transcription in vivo: the aromatic hydrocarbon receptor mediates transactivation, enhancer-promoter communication, and changes in chromatin structure. Mol Cell Biol. 1996 Jan;16(1):430–436. doi: 10.1128/mcb.16.1.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kotani H., Newton P. B., 3rd, Zhang S., Chiang Y. L., Otto E., Weaver L., Blaese R. M., Anderson W. F., McGarrity G. J. Improved methods of retroviral vector transduction and production for gene therapy. Hum Gene Ther. 1994 Jan;5(1):19–28. doi: 10.1089/hum.1994.5.1-19. [DOI] [PubMed] [Google Scholar]
  34. Li H., Dong L., Whitlock J. P., Jr Transcriptional activation function of the mouse Ah receptor nuclear translocator. J Biol Chem. 1994 Nov 11;269(45):28098–28105. [PubMed] [Google Scholar]
  35. Lin Y. S., Green M. R. Mechanism of action of an acidic transcriptional activator in vitro. Cell. 1991 Mar 8;64(5):971–981. doi: 10.1016/0092-8674(91)90321-o. [DOI] [PubMed] [Google Scholar]
  36. Ma Q., Dong L., Whitlock J. P., Jr Transcriptional activation by the mouse Ah receptor. Interplay between multiple stimulatory and inhibitory functions. J Biol Chem. 1995 May 26;270(21):12697–12703. doi: 10.1074/jbc.270.21.12697. [DOI] [PubMed] [Google Scholar]
  37. Miller A. G., Israel D., Whitlock J. P., Jr Biochemical and genetic analysis of variant mouse hepatoma cells defective in the induction of benzo(a)pyrene-metabolizing enzyme activity. J Biol Chem. 1983 Mar 25;258(6):3523–3527. [PubMed] [Google Scholar]
  38. Mizzen C. A., Yang X. J., Kokubo T., Brownell J. E., Bannister A. J., Owen-Hughes T., Workman J., Wang L., Berger S. L., Kouzarides T. The TAF(II)250 subunit of TFIID has histone acetyltransferase activity. Cell. 1996 Dec 27;87(7):1261–1270. doi: 10.1016/s0092-8674(00)81821-8. [DOI] [PubMed] [Google Scholar]
  39. Morgan J. E., Whitlock J. P., Jr Transcription-dependent and transcription-independent nucleosome disruption induced by dioxin. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11622–11626. doi: 10.1073/pnas.89.23.11622. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Nebert D. W., Gonzalez F. J. P450 genes: structure, evolution, and regulation. Annu Rev Biochem. 1987;56:945–993. doi: 10.1146/annurev.bi.56.070187.004501. [DOI] [PubMed] [Google Scholar]
  41. Nelson R. M., Long G. L. A general method of site-specific mutagenesis using a modification of the Thermus aquaticus polymerase chain reaction. Anal Biochem. 1989 Jul;180(1):147–151. doi: 10.1016/0003-2697(89)90103-6. [DOI] [PubMed] [Google Scholar]
  42. Okey A. B., Riddick D. S., Harper P. A. Molecular biology of the aromatic hydrocarbon (dioxin) receptor. Trends Pharmacol Sci. 1994 Jul;15(7):226–232. doi: 10.1016/0165-6147(94)90316-6. [DOI] [PubMed] [Google Scholar]
  43. Okino S. T., Whitlock J. P., Jr Dioxin induces localized, graded changes in chromatin structure: implications for Cyp1A1 gene transcription. Mol Cell Biol. 1995 Jul;15(7):3714–3721. doi: 10.1128/mcb.15.7.3714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Pear W. S., Nolan G. P., Scott M. L., Baltimore D. Production of high-titer helper-free retroviruses by transient transfection. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8392–8396. doi: 10.1073/pnas.90.18.8392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Peterson C. L., Tamkun J. W. The SWI-SNF complex: a chromatin remodeling machine? Trends Biochem Sci. 1995 Apr;20(4):143–146. doi: 10.1016/s0968-0004(00)88990-2. [DOI] [PubMed] [Google Scholar]
  46. Poellinger L., Göttlicher M., Gustafsson J. A. The dioxin and peroxisome proliferator-activated receptors: nuclear receptors in search of endogenous ligands. Trends Pharmacol Sci. 1992 Jun;13(6):241–245. doi: 10.1016/0165-6147(92)90076-i. [DOI] [PubMed] [Google Scholar]
  47. Poland A., Knutson J. C. 2,3,7,8-tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons: examination of the mechanism of toxicity. Annu Rev Pharmacol Toxicol. 1982;22:517–554. doi: 10.1146/annurev.pa.22.040182.002505. [DOI] [PubMed] [Google Scholar]
  48. Reisz-Porszasz S., Probst M. R., Fukunaga B. N., Hankinson O. Identification of functional domains of the aryl hydrocarbon receptor nuclear translocator protein (ARNT). Mol Cell Biol. 1994 Sep;14(9):6075–6086. doi: 10.1128/mcb.14.9.6075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Roberts S. G., Ha I., Maldonado E., Reinberg D., Green M. R. Interaction between an acidic activator and transcription factor TFIIB is required for transcriptional activation. Nature. 1993 Jun 24;363(6431):741–744. doi: 10.1038/363741a0. [DOI] [PubMed] [Google Scholar]
  50. Rowlands J. C., McEwan I. J., Gustafsson J. A. Trans-activation by the human aryl hydrocarbon receptor and aryl hydrocarbon receptor nuclear translocator proteins: direct interactions with basal transcription factors. Mol Pharmacol. 1996 Sep;50(3):538–548. [PubMed] [Google Scholar]
  51. Safe S. H. Comparative toxicology and mechanism of action of polychlorinated dibenzo-p-dioxins and dibenzofurans. Annu Rev Pharmacol Toxicol. 1986;26:371–399. doi: 10.1146/annurev.pa.26.040186.002103. [DOI] [PubMed] [Google Scholar]
  52. Sogawa K., Iwabuchi K., Abe H., Fujii-Kuriyama Y. Transcriptional activation domains of the Ah receptor and Ah receptor nuclear translocator. J Cancer Res Clin Oncol. 1995;121(9-10):612–620. doi: 10.1007/BF01197779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Sogawa K., Nakano R., Kobayashi A., Kikuchi Y., Ohe N., Matsushita N., Fujii-Kuriyama Y. Possible function of Ah receptor nuclear translocator (Arnt) homodimer in transcriptional regulation. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):1936–1940. doi: 10.1073/pnas.92.6.1936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Stringer K. F., Ingles C. J., Greenblatt J. Direct and selective binding of an acidic transcriptional activation domain to the TATA-box factor TFIID. Nature. 1990 Jun 28;345(6278):783–786. doi: 10.1038/345783a0. [DOI] [PubMed] [Google Scholar]
  55. Struhl K. Chromatin structure and RNA polymerase II connection: implications for transcription. Cell. 1996 Jan 26;84(2):179–182. doi: 10.1016/s0092-8674(00)80970-8. [DOI] [PubMed] [Google Scholar]
  56. Swanson H. I., Bradfield C. A. The AH-receptor: genetics, structure and function. Pharmacogenetics. 1993 Oct;3(5):213–230. doi: 10.1097/00008571-199310000-00001. [DOI] [PubMed] [Google Scholar]
  57. Triezenberg S. J. Structure and function of transcriptional activation domains. Curr Opin Genet Dev. 1995 Apr;5(2):190–196. doi: 10.1016/0959-437x(95)80007-7. [DOI] [PubMed] [Google Scholar]
  58. Whitelaw M. L., Gustafsson J. A., Poellinger L. Identification of transactivation and repression functions of the dioxin receptor and its basic helix-loop-helix/PAS partner factor Arnt: inducible versus constitutive modes of regulation. Mol Cell Biol. 1994 Dec;14(12):8343–8355. doi: 10.1128/mcb.14.12.8343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Whitlock J. P., Jr Mechanistic aspects of dioxin action. Chem Res Toxicol. 1993 Nov-Dec;6(6):754–763. doi: 10.1021/tx00036a003. [DOI] [PubMed] [Google Scholar]
  60. Whitlock J. P., Jr, Okino S. T., Dong L., Ko H. P., Clarke-Katzenberg R., Ma Q., Li H. Cytochromes P450 5: induction of cytochrome P4501A1: a model for analyzing mammalian gene transcription. FASEB J. 1996 Jun;10(8):809–818. doi: 10.1096/fasebj.10.8.8666157. [DOI] [PubMed] [Google Scholar]
  61. Wilson C. J., Chao D. M., Imbalzano A. N., Schnitzler G. R., Kingston R. E., Young R. A. RNA polymerase II holoenzyme contains SWI/SNF regulators involved in chromatin remodeling. Cell. 1996 Jan 26;84(2):235–244. doi: 10.1016/s0092-8674(00)80978-2. [DOI] [PubMed] [Google Scholar]
  62. Wolffe A. P., Pruss D. Targeting chromatin disruption: Transcription regulators that acetylate histones. Cell. 1996 Mar 22;84(6):817–819. doi: 10.1016/s0092-8674(00)81059-4. [DOI] [PubMed] [Google Scholar]
  63. Workman J. L., Taylor I. C., Kingston R. E. Activation domains of stably bound GAL4 derivatives alleviate repression of promoters by nucleosomes. Cell. 1991 Feb 8;64(3):533–544. doi: 10.1016/0092-8674(91)90237-s. [DOI] [PubMed] [Google Scholar]
  64. Wu L., Whitlock J. P., Jr Mechanism of dioxin action: Ah receptor-mediated increase in promoter accessibility in vivo. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4811–4815. doi: 10.1073/pnas.89.11.4811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Xiao H., Pearson A., Coulombe B., Truant R., Zhang S., Regier J. L., Triezenberg S. J., Reinberg D., Flores O., Ingles C. J. Binding of basal transcription factor TFIIH to the acidic activation domains of VP16 and p53. Mol Cell Biol. 1994 Oct;14(10):7013–7024. doi: 10.1128/mcb.14.10.7013. [DOI] [PMC free article] [PubMed] [Google Scholar]

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