The brown shrimp (Crangon crangon L.) ecdysteroid receptor complex: cloning, structural modeling of the ligand-binding domain and functional expression in an EcR-deficient Drosophila cell line.
cDNAs encoding ecdysteroid receptor (EcR) and retinoid X receptor (RXR) were cloned and sequenced from brown shrimp Crangon crangon (Crustacea: Decapoda), a common faunal species and commercially important in the North-West European coastal waters. A 3D model of the ligand-binding domain (LBD) of EcR was created and docking of ponasterone A (PonA) was simulated in silico. Finally, we report the transfection of expression plasmids for these receptors in the mutant Drosophila L57-3-11 cell line. Through an ecdysteroid responsive reporter assay we clearly prove the functionality of shrimp ecdysteroid receptor in the transfected L57-3-11 cell line. Our results indicate that the Drosophila L57-3-11 cell line and in silico LBD modeling can be used to study the function of crustacean ecdysteroid receptors and be applied to assess endocrine disrupting effects on non-target crustacean species.
Laboratory of Agrozoology, Faculty of Bioscience Engineering, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium.
Yves
Y
Verhaegen
Koen
K
Parmentier
Luc
L
Swevers
Pierre
P
Rougé
Thomas
T
Soin
Wim
W
De Coen
Kris
K
Cooreman
Guy
G
Smagghe
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D033501:Crangonidae / Q000378:metabolism*; D004330:Drosophila; D004441:Ecdysterone / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D017433:Protein Structure, Secondary; D011987:Receptors, Steroid / Q000737:chemistry* / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D047488:Retinoid X Receptors / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D011987:Receptors, Steroid; D047488:Retinoid X Receptors; C034808:ecdysteroid receptor; D004441:Ecdysterone; C008999:ponasterone A