title: Structural features critical to the activity of an ecdysone receptor binding site.
abstract: Two ecdysone-response elements from the hsp27 (hsp27 EcRE) and the Fbp1 (D EcRE) genes of Drosophila melanogaster were used as probes in a gel shift assay to investigate the interactions of the ecdysone receptor (EcR) with its cognate DNA response element. The source of EcR was a nuclear extract from the late third-larval instar fat body. The hsp27 and D EcREs share a sequence similarity at 12 positions over a 15bp region including an imperfect palindromic structure consisting of two pentamer half-sites separated by a single intervening nucleotide. We have shown that a short oligonucleotide containing this 11bp imperfect palindrome of the hsp27 EcRE and three flanking bp on each side is an efficient EcR binding site. Mutational analysis confirms that the integrity of both these half-sites as well as their 1bp spacing are critical for binding of the ecdysone receptor. The D EcRE behaved as a much weaker EcR binding site than the hsp27 EcRE but a single bp substitution was sufficient to confer upon it a binding capacity equivalent to that of the hsp27 EcRE. These results have led us to propose the sequence PuG(G/T)T(C/G)A(N)TG(C/A)(C/A)(C/t)Py as a revised version of a previously proposed EcRE consensus sequence.
authors: Institut Jacques Monod, CNRS, Paris, France.
authors: C
authors: C
authors: Antoniewski
authors: M
authors: M
authors: Laval
authors: J A
authors: JA
authors: Lepesant
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics / Q000378:metabolism; D015342:DNA Probes; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D004440:Ecdysone / Q000378:metabolism*; D005216:Fat Body / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid
chemical_list: D015342:DNA Probes; D011987:Receptors, Steroid; C055469:ecdysone receptor; D004440:Ecdysone; D004247:DNA