Skip to main content
Springer Nature Link
Log in

Aryl hydrocarbon receptor (AhR)-mediated reporter gene expression systems in transgenic tobacco plants

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

In mammals, the aryl hydrocarbon receptor (AhR) mediates expression of certain genes, including CYP1A1, in response to exposure to dioxins and related compounds. We have constructed a mouse AhR-mediated gene expression systems for a β-glucuronidase (GUS) reporter gene consisting of an AhR, an AhR nuclear translocator (Arnt), and a xenobiotic response element (XRE)-driven promoter in transgenic tobacco plants. On treatment with the AhR ligands 3-methylcholanthrene (MC), β-naphthoflavone (βNF), and indigo, the transgenic tobacco plants exhibited enhanced GUS activity, presumably by inducible expression of the reporter gene. The recombinant AhR (AhRV), with the activation domain replaced by that of the Herpes simplex virus protein VP16, induced GUS activity much more than the wild-type AhR in the transgenic tobacco plants. Plants carrying AhRV expressed the GUS reporter gene in a dose- and time-dependent manner when treated with MC; GUS activity was detected at 5 nM MC on solid medium and at 12 h after soaking in 25 μM MC. Histochemical GUS staining showed that this system was active mainly in leaf and stem. These results suggest that the AhR-mediated reporter gene expression system has potential for the bioassay of dioxins in the environment and as a novel gene expression system in plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

Abbreviations

AhR:

Aryl hydrocarbon receptor

Arnt:

AhR nuclear translocator

CYP1A1:

Cytochrome P450 1A1

GUS:

β-Glucuronidase

Hsp90:

90-kDa Heat shock protein

IMM:

Immunophilin-like protein

MC:

3-Methylcholanthrene

MS:

Murashige and Skoog

4MU:

4-Methyl-umbelliferone

4MUG:

4-Methyl-umbelliferyl-β-d-glucuronide

βNF:

β-Naphthoflavone

Nos-T:

Nos terminator

PCB:

Polychlorinated biphenyl

PCDF:

Polychlorinated dibenzofuran

PCDD:

Polychlorinated dibenzo-p-dioxin

POP:

Persistent organic pollutant

35-S:

CaMV35S promoter

35Sm-P:

−60/+8 minimal CaMV35S promoter

TAD:

Transactivation domain

X-Gluc:

5-Bromo-4-chloro-3-indolyl-β-d-glucuronide

XRE:

Xenobiotic respons element

XRE-P:

XRE-driven promoter

References

  • Anderson JW, Hartwell SI, Hameedi MJ (2005) Regional comparisons of coastal sediment contamination detected by a biomarker (P450 HRGS; EPA Method 4425). Environ Sci Technol 39:17–23

    Article  PubMed  CAS  Google Scholar 

  • Aoyama T, Chua NH (1997) A glucocorticoid-mediated transcriptional induction system in transgenic plants. Plant J 11:605–612

    Article  PubMed  CAS  Google Scholar 

  • Bank PA, Yao EF, Phelps CL, Harper PA, Denison MS (1992) Species-specific binding of transformed Ah receptor to a dioxin responsive transcriptional enhancer. Eur J Pharmacol 228:85–94

    PubMed  CAS  Google Scholar 

  • Carver LA, Jackiw V, Bradfield CA (1994) The 90-kDa heat shock protein is essential for Ah receptor signaling in a yeast expression system. J Biol Chem 269:30109–30112

    PubMed  CAS  Google Scholar 

  • Clark G, Tritscher A, Bell D, Lucier G (1992) Integrated approach for evaluating species and interindividual differences in responsiveness to dioxins and structural analogs. Environ Health Perspect 98:125–132

    Article  PubMed  CAS  Google Scholar 

  • Dalrymple MA, McGeoch DJ, Davison AJ, Preston CM (1985) DNA sequence of the herpes simplex virus type 1 gene whose product is responsible for transcriptional activation of immediate early promoters. Nucleic Acids Res 13:7865–7879

    Article  PubMed  CAS  Google Scholar 

  • Ema M, Sogawa K, Watanabe N, Chujoh Y, Matsushita N, Gotoh O, Funae Y, Fujii-Kuriyama Y (1992) cDNA cloning and structure of mouse putative Ah receptor. Biochem Biophys Res Commun 184:246–253

    Article  PubMed  CAS  Google Scholar 

  • Ema M, Ohe N, Suzuki M, Mimura J, Sogawa K, Ikawa S, Fujii-Kuriyama Y (1994) Dioxin binding activities of polymorphic forms of mouse and human arylhydrocarbon receptors. J Biol Chem 269:27337–27343

    PubMed  CAS  Google Scholar 

  • Harrell JM, Kurek I, Breiman A, Radanyi C, Renoir JM, Pratt WB, Galigniana MD (2002) All of the protein interactions that link steroid receptor·hsp90·immunophilin heterocomplexes to cytoplasmic dynein are common to plant and animal cells. Biochemistry 41:5581–5587

    Article  PubMed  CAS  Google Scholar 

  • Harvey PJ, Campanella BF, Castro PM, Harms H, Lichtfouse E, Schaffner AR, Smrcek S, Werck-Reichhart D (2002) Phytoremediation of polyaromatic hydrocarbons, anilines and phenols. Environ Sci Pollut Res Int 9:29–47

    Article  PubMed  CAS  Google Scholar 

  • Hoivik DJ, Safe SH, Gaido KW (1998) Effects of xenobiotics on hormone receptors. Taylor & Francis, Bristol, pp 53–68

    Google Scholar 

  • Hollingshead BD, Petrulis JR, Perdew GH (2004) The aryl hydrocarbon (Ah) receptor transcriptional regulator hepatitis B virus X-associated protein 2 antagonizes p23 binding to Ah receptor-Hsp90 complexes and is dispensable for receptor function. J Biol Chem 279:45652–45661

    Article  PubMed  CAS  Google Scholar 

  • Horsch RB, Fry JE, Hoffmann NL, Eichholz D, Rogers DG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231

    Article  CAS  Google Scholar 

  • Hülster A, Müller JF, Marschner H (1994) Soil-plant transfer of polychlorinated dibenzo-p-dioxins and dibenzofurans to vegetables of the cucumber family. Environ Sci Technol 28:1110–1115

    Article  Google Scholar 

  • Inui H, Kodama T, Ohkawa Y, Ohkawa H (2000) Herbicide metabolism and cross-tolerance in transgenic potato plants co-expressing human CYP1A1, CYP2B6 and CYP2C19. Pestic Biochem Physiol 66:116–129

    Article  CAS  Google Scholar 

  • Inui H, Sasaki H, Kodama S, Chua N-H, Ohkawa H (2005) Monitoring of endocrine disruptors in transgenic plants carrying aryl hydrocarbon receptor and estrogen receptor genes. In: Clark JM, Ohkawa H (eds) New discoveries in agrochemicals. American Chemical Society, Washington DC, pp 40–47

    Google Scholar 

  • Kazlauskas A, Poellinger L, Pongratz I (1999) Evidence that the co-chaperone p23 regulates ligand responsiveness of the dioxin (Aryl hydrocarbon) receptor. J Biol Chem 274:13519–13524

    Article  PubMed  CAS  Google Scholar 

  • Kazlauskas A, Sundstrom S, Poellinger L, Pongratz I (2001) The hsp90 chaperone complex regulates intracellular localization of the dioxin receptor. Mol Cell Biol 21:2594–2607

    Article  PubMed  CAS  Google Scholar 

  • Ko HP, Okino ST, Ma Q, Whitlock JP Jr (1996) Dioxin-induced CYP1A1 transcription in vivo: the aromatic hydrocarbon receptor mediates transactivation, enhancer-promoter communication, and changes in chromatin structure. Mol Cell Biol 16:430–436

    PubMed  CAS  Google Scholar 

  • Korkalainen M, Tuomisto J, Pohjanvirta R (2001) The AH receptor of the most dioxin-sensitive species, guinea pig, is highly homologous to the human AH receptor. Biochem Biophys Res Commun 285:1121–1129

    Article  PubMed  CAS  Google Scholar 

  • Lees MJ, Whitelaw ML (1999) Multiple roles of ligand in transforming the dioxin receptor to an active basic helix-loop-helix/PAS transcription factor complex with the nuclear protein Arnt. Mol Cell Biol 19:5811–5822

    PubMed  CAS  Google Scholar 

  • Lin YS, Ha I, Maldonado E, Reinberg D, Green MR (1991) Binding of general transcription factor TFIIB to an acidic activating region. Nature 353:569–571

    Article  PubMed  CAS  Google Scholar 

  • Lusska A, Shen E, Whitlock JP Jr (1993) Protein-DNA interactions at a dioxin-responsive enhancer. Analysis of six bona fide DNA-binding sites for the liganded Ah receptor. J Biol Chem 268:6575–6580

    PubMed  CAS  Google Scholar 

  • Martinez A, Sparks C, Drayton P, Thompson J, Greenland A, Jepson I (1999a) Creation of ecdysone receptor chimeras in plants for controlled regulation of gene expression. Mol Gen Genet 261:546–552

    Article  PubMed  CAS  Google Scholar 

  • Martinez A, Sparks C, Hart CA, Thompson J, Jepson I (1999b) Ecdysone agonist inducible transcription in transgenic tobacco plants. Plant J 19:97–106

    Article  PubMed  CAS  Google Scholar 

  • Miller CA 3rd (1997) Expression of the human aryl hydrocarbon receptor complex in yeast. Activation of transcription by indole compounds. J Biol Chem 272:32824–32829

    Article  PubMed  CAS  Google Scholar 

  • Picard D (2006) Chaperoning steroid hormone action. Trends Endocrinol Metab 17:229–235

    Article  PubMed  CAS  Google Scholar 

  • Reisz-Porszasz S, Probst MR, Fukunaga BN, Hankinson O (1994) Identification of functional domains of the aryl hydrocarbon receptor nuclear translocator protein (ARNT). Mol Cell Biol 14:6075–6086

    PubMed  CAS  Google Scholar 

  • Rowlands JC, Gustafsson JA (1997) Aryl hydrocarbon receptor-mediated signal transduction. Crit Rev Toxicol 27:109–134

    Article  PubMed  CAS  Google Scholar 

  • Sakaki T, Shinkyo R, Takita T, Ohta M, Inouye K (2002) Biodegradation of polychlorinated dibenzo-p-dioxins by recombinant yeast expressing rat CYP1A subfamily. Arch Biochem Biophys 401:91–98

    Article  PubMed  CAS  Google Scholar 

  • Schwechheimer C, Smith C, Bevan MW (1998) The activities of acidic and glutamine-rich transcriptional activation domains in plant cells: design of modular transcription factors for high-level expression. Plant Mol Biol 36:195–204

    Article  PubMed  CAS  Google Scholar 

  • Shen F, Triezenberg SJ, Hensley P, Porter D, Knutson JR (1996) Transcriptional activation domain of the herpesvirus protein VP16 becomes conformationally constrained upon interaction with basal transcription factors. J Biol Chem 271:4827–4837

    Article  PubMed  CAS  Google Scholar 

  • Stancato LF, Hutchison KA, Krishna P, Pratt WB (1996) Animal and plant cell lysates share a conserved chaperone system that assembles the glucocorticoid receptor into a functional heterocomplex with hsp90. Biochemistry 35:554–561

    Article  PubMed  CAS  Google Scholar 

  • Thangavelu M, Belostotsky D, Bevan MW, Flavell RB, Rogers HJ, Lonsdale DM (1993) Partial characterization of the Nicotiana tabacum actin gene family: evidence for pollen-specific expression of one of the gene family members. Mol Gen Genet 240:290–295

    Article  PubMed  CAS  Google Scholar 

  • Tojo T, Tsuda K, Wada TS, Yamazaki K (2006) A simple and extremely sensitive system for detecting estrogenic activity using transgenic Arabidopsis thaliana. Ecotoxicol Environ Saf 64:106–114

    Article  PubMed  CAS  Google Scholar 

  • Whitelaw ML, McGuire J, Picard D, Gustafsson JA, Poellinger L (1995) Heat shock protein hsp90 regulates dioxin receptor function in vivo. Proc Natl Acad Sci USA 92:4437–4441

    Article  PubMed  CAS  Google Scholar 

  • Whitlock JP Jr (1999) Induction of cytochrome P4501A1. Annu Rev Pharmacol Toxicol 39:103–125

    Article  PubMed  CAS  Google Scholar 

  • Windal I, Denison MS, Birnbaum LS, Van Wouwe N, Baeyens W, Goeyens L (2005) Chemically activated luciferase gene expression (CALUX) cell bioassay analysis for the estimation of dioxin-like activity: critical parameters of the CALUX procedure that impact assay results. Environ Sci Technol 39:7357–7364

    Article  PubMed  CAS  Google Scholar 

  • Wong JM, Okey AB, Harper PA (2001) Human aryl hydrocarbon receptor polymorphisms that result in loss of CYP1A1 induction. Biochem Biophys Res Commun 288:990–996

    Article  PubMed  CAS  Google Scholar 

  • Zuo J, Niu QW, Chua N-H (2000) Technical advance: an estrogen receptor-based transactivator XVE mediates highly inducible gene expression in transgenic plants. Plant J 24:265–273

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the Bio-oriented Technology Research Advancement Institution (BRAIN), Tokyo, Japan.

Author information

Author notes

  1. Susumu Kodama

    Present address: Pharmacogenetics Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC, 27709, USA

  2. Hideo Ohkawa

    Present address: Research Center for Green Science, Fukuyama University, Gakuen-cho 1, Fukuyama, Hiroshima, 729-0292, Japan

Authors and Affiliations

  1. Graduate School of Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, Hyogo, 657-8501, Japan

    Susumu Kodama & Kumiko Okada

  2. Research Center for Environmental Genomics, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, Hyogo, 657-8501, Japan

    Hideyuki Inui & Hideo Ohkawa

Authors
  1. Susumu Kodama
    View author publications

    Search author on:PubMed Google Scholar

  2. Kumiko Okada
    View author publications

    Search author on:PubMed Google Scholar

  3. Hideyuki Inui
    View author publications

    Search author on:PubMed Google Scholar

  4. Hideo Ohkawa
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Hideo Ohkawa.

Electronic supplementary material

Below is the link to the electronic supplementary material.

425_2007_592_MOESM1_ESM.pdf

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kodama, S., Okada, K., Inui, H. et al. Aryl hydrocarbon receptor (AhR)-mediated reporter gene expression systems in transgenic tobacco plants. Planta 227, 37–45 (2007). https://doi.org/10.1007/s00425-007-0592-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-007-0592-1

Keywords

Profiles

  1. Hideyuki Inui View author profile