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

Gen Comp Endocrinol. 2010 Sep 15;168(3):415-23. doi: 10.1016/j.ygcen.2010.05.007. Epub 2010 May 31.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Line
  • Crangonidae / metabolism*
  • Drosophila
  • Ecdysterone / analogs & derivatives
  • Ecdysterone / chemistry
  • Ecdysterone / metabolism
  • Molecular Sequence Data
  • Phylogeny
  • Protein Structure, Secondary
  • Receptors, Steroid / chemistry*
  • Receptors, Steroid / classification
  • Receptors, Steroid / genetics
  • Receptors, Steroid / metabolism*
  • Retinoid X Receptors / chemistry
  • Retinoid X Receptors / metabolism
  • Sequence Homology, Amino Acid

Substances

  • Receptors, Steroid
  • Retinoid X Receptors
  • ecdysteroid receptor
  • Ecdysterone
  • ponasterone A