The heterodimeric ecdysteroid receptor complex in the brown shrimp Crangon crangon: EcR and RXR isoform characteristics and sensitivity towards the marine pollutant tributyltin |
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Authors: | Verhaegen Yves Parmentier Koen Swevers Luc Renders Ellen Rougé Pierre De Coen Wim Cooreman Kris Smagghe Guy |
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Affiliation: | a Laboratory of Agrozoology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium b Institute for Agricultural and Fisheries Research, Animal Sciences Unit, Fisheries Department, Ankerstraat 1, B-8400 Ostend, Belgium c Laboratory of Ecophysiology, Biochemistry and Toxicology, Department of Biology, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp, Belgium d Insect Molecular Genetics and Biotechnology, Institute of Biology, National Centre for Scientific Research “Demokritos”, Aghia Paraskevi Attikis, Athens, Greece e Surfaces Cellulaires et Signalisation chez les Végétaux, UMR Université Paul Sabatier CNRS 5546, Castanet Tolosan, France |
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Abstract: | Decapod crustaceans are characterized by multiple ecdysteroid receptor (EcR) and retinoid-X-receptor (RXR) isoforms, which likely exhibit variant dimerization and transactivation interactions. In the brown shrimp C. crangon we cloned C-terminally truncated CrcEcR and CrcRXR isoforms and isoforms exhibiting deletions within the hinge region. For the former, in silico modeling of the CrcEcR indicated that, where the conserved helices H10 and H11 of the ligand-binding domain (LBD) are missing, an alternative C-terminal α-helix repairs the ligand-binding pocket (LBP). The truncated CrcRXR isoforms lack a major part of the LBD (H4-H12), thereby compromising ligand binding and dimerization. Through an in vitro ecdysteroid responsive reporter assay, we showed that these natural receptor variations do not impair receptor functioning but probably alter the receptor dimerization preferences. By the same in vitro assay, using full-length CrcEcR and CrcRXR, the effect of tributyltin (TBT) on ecdysteroid-induced transactivation was evaluated. The transactivation by 10 nM PonA was reduced with 64% by 20 nM TBT. In silico modeling confirmed that TBT fits in the full-length CrcRXR-LBD. Furthermore, semi-quantitative PCR indicated altered expression of CrcEcR and CrcRXR isoforms after in vivo acute exposure to TBT, especially in the ovaries. |
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Keywords: | Crangon crangon Shrimp Crustacea Tributyltin TBT Retinoid-X-receptor RXR Ecdysteroid receptor EcR |
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