Characterization of the region of the aryl hydrocarbon receptor required for ligand dependency of transactivation using chimeric receptor between Drosophila and Mus musculus
Abstract
The aryl hydrocarbon receptor (AhR) is a ligand-activated transcriptional factor. Although 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is high affinity and toxic to many vertebrate animals, invertebrate AhRs including Drosophila melanogaster AhR (spineless) have no ability to bind exogenous chemicals as ligands. To analyze the ligand-binding domain (LBD) of AhR, we used chimeras between mouse and Drosophila AhR. The chimeric AhR revealed that the LBD determines constitutive transactivation in Drosophila AhR or ligand-dependent activation in mouse AhR. The LBD was further divided into three blocks that corresponded to amino acids 230-300, 301-361, and 361-420 of the mouse sequence. Six chimeric proteins clarified that amino acids 291-350 of the Drosophila LBD, i.e. the middle region, were required to keep the protein in the active form in the absence of ligand binding, whereas in the mouse AhR, this region was required to maintain the protein in the inactive form in the absence of ligand. Furthermore, Arg346 in the middle region of the mouse LBD, was identified as amino acids that were critical for AhR activation by site-directed mutagenesis.
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A prior investigation employing chimeric AHR proteins from mouse and drosophila demonstrated that the middle region of the AHR PAS-B domain governs ligand responsiveness in transactivation.
“Intriguingly, a prior investigation employing chimeric AHR proteins from both mouse and drosophila demonstrated that the middle region of the AHR PAS-B domain governs ligand responsiveness in transactivation47 (Supplementary Fig. 9f).”
From Structural basis for the ligand-dependent activation of heterodimeric AHR-ARNT complex (2025) · cites this paper as [47]
dAHR is known to be constitutively active in a ligand-independent manner.
“We then expanded our structural comparison to the dAHR, which is known to be constitutively active in a ligand-independent manner47.”
From Structural basis for the ligand-dependent activation of heterodimeric AHR-ARNT complex (2025) · cites this paper as [47]
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Zhao, H., Tan, J., Tang, T., & Dai, S. (2026) · TrAC Trends in Analytical Chemistry · doi:10.1016/j.trac.2026.118932
- Authors
- Kudo, K., Takeuchi, T., Murakami, Y., Ebina, M., & Kikuchi, H.
- Affiliations
- Iwate University (JP)
- Tohoku University (JP)
- Hirosaki University (JP)
- Corresponding author
- Kikuchi, Hideaki
- Venue
- Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms
- Publisher
- Elsevier BV
- Published
- 2009-06
- Volume
- 1789
- Issue
- 6-8
- Pages
- 477-486
- ISSN
- 1874-9399 (ISSN-L)
- OpenAlex
- W2102844036
- Cited by
- 25
- References
- 36
- Citations by year
- 2026: 3 · 2025: 4 · 2023: 3 · 2022: 2 · 2020: 2 · 2017: 1 · 2014: 2 · 2012: 2
- FWCI
- 1.89
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Toxic Organic Pollutants Impact
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
- Keywords
- aryl hydrocarbon receptor, ligand-binding domain, transcriptional activation, chimeric receptors, Drosophila melanogaster, mice, tetrachlorodibenzo-p-dioxin, constitutive activation, ligand-dependent activation, transactivation domain
- MeSH terms
- Polychlorinated Dibenzodioxins, Amino Acid Sequence, Animals, Base Sequence, Binding Sites, Cell Line, Ligands, Models, Molecular, Molecular Sequence Data, Recombinant Fusion Proteins, Species Specificity, Transcriptional Activation, Mutagenesis, Site-Directed, Sequence Homology, Amino Acid, Protein Structure, Tertiary, DNA Primers, Receptors, Aryl Hydrocarbon, Amino Acid Substitution, Drosophila Proteins, Mice
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