Citation

  • Authors: Maletta, M., Orlov, I., Roblin, P., Beck, Y., Moras, D., Billas, I. M., Klaholz, B. P.
  • Year: 2014
  • Journal: Nat Commun 5 4139
  • Applications: in vitro / DNA / jetPEI
  • Cell types:
    1. Name: COS-1
      Description: African green monkey kidney cells
      Known as: COS, COS1
    2. Name: HEK 293 EBNA
      Description: Transformed with adenovirus 5 DNA Increased recombinant protein yield with EBV oriP vector
      Known as: 293 c18 ; 293 EBNA ; ATCC® CRL-10852™
    3. Name: HEK-293
      Description: Human embryonic kidney Fibroblast
      Known as: HEK293, 293

Abstract

Nuclear receptors (NRs) regulate gene expression through DNA- and ligand-binding and thus represent crucial therapeutic targets. The ultraspiracle protein/ecdysone receptor (USP/EcR) complex binds to half-sites with a one base pair spaced inverted repeat (IR1), a palindromic DNA response element (RE) reminiscent of IRs observed for vertebrate steroid hormone receptors. Here we present the cryo electron microscopy structure of the USP/EcR complex bound to an IR1 RE which provides the first description of a full IR-bound NR complex. The structure reveals that even though the DNA is almost symmetric, the complex adopts a highly asymmetric architecture in which the ligand-binding domains (LBDs) are positioned 5' off-centred. Additional interactions of the USP LBD with the 5'-flanking sequence trigger transcription activity as monitored by transfection assays. The comparison with DR-bound NR complexes suggests that DNA is the major allosteric driver in inversely positioning the LBDs, which serve as the main binding-site for transcriptional regulators.

Go to