HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 26, 27211-27218, June 25, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/26/27211    most recent M403839200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kumar, M. B. Articles by Palli, S. R. Search for Related Content PubMed PubMed Citation Articles by Kumar, M. B. Articles by Palli, S. R. Social Bookmarking                      What's this?

Highly Flexible Ligand Binding Pocket of Ecdysone Receptor

A SINGLE AMINO ACID CHANGE LEADS TO DISCRIMINATION BETWEEN TWO GROUPS OF NONSTEROIDAL ECDYSONE AGONISTS^*

Mohan B. Kumar, David W. Potter, Robert E. Hormann, Angela Edwards, Colin M. Tice, Howard C. Smith, Martha A. Dipietro, Mitch Polley, Michael Lawless, Philippa R. N. Wolohan, Damodhar R. Kethidi¶, and Subba R. Palli¶||

From the RheoGene Inc., Norristown, Pennsylvania 19403, Tripos Inc., St. Louis, Missouri 63144, and the ^¶Department of Entomology, College of Agriculture, University of Kentucky, Lexington, Kentucky 40546

The insect steroid hormone 20-hydroxyecdysone works through a ligand-activated nuclear receptor, the ecdysone receptor (EcR), which plays critical roles in insect development and reproduction. The EcR has been exploited to develop insecticides to control pests and gene switches for gene regulation. Recently reported crystal structures of the EcR protein show different but partially overlapping binding cavities for ecdysteroid (ECD) and diacylhydrazine (DAH) ligands, providing an explanation for the differential activity of DAH ligands in insects. 1-Aroyl-4-(arylamino)-1,2,3,4-tetrahydroquinoline (THQ) ligands were recently discovered as ecdysone agonists. Mutagenesis of the EcR (from Choristoneura fumiferana, CfEcR) ligand binding domain followed by screening in a reporter assay led to the identification of CfEcR mutants, which responded well to THQ ligands but poorly to both ECD and DAH ligands. These mutants were further improved by introducing a second mutation, A110P, which was previously reported to cause ECD insensitivity. Testing of these V128F/A110P and V128Y/A110P mutants in a C57BL/6 mouse model coactivator interaction assay and in insect cells showed that this mutant EcR is activated by THQ ligands but not by ECD or DAH ligands. The CfEcR and its V128F/A110P mutant were used to demonstrate simultaneous regulation of two reporter genes using THQ and DAH ligands.

Received for publication, April 6, 2004 , and in revised form, April 22, 2004.

^* This work was supported in part by National Institute of Standards and Technology Advanced Technology Project Grant 70NANB0H3012 to RheoGene Inc. and a RheoGene Inc. research grant (to S. R. P.). This is contribution number 04-08-075 from the Kentucky Agricultural Experimental Station. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Dept. of Entomology, College of Agriculture, University of Kentucky, Lexington, KY 40546. Tel.: 859-257-4962; Fax: 859-323-1120; Email: rpalli{at}uky.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. A. Carmichael, M. C. Lawrence, L. D. Graham, P. A. Pilling, V. C. Epa, L. Noyce, G. Lovrecz, D. A. Winkler, A. Pawlak-Skrzecz, R. E. Eaton, et al. The X-ray Structure of a Hemipteran Ecdysone Receptor Ligand-binding Domain: COMPARISON WITH A LEPIDOPTERAN ECDYSONE RECEPTOR LIGAND-BINDING DOMAIN AND IMPLICATIONS FOR INSECTICIDE DESIGN J. Biol. Chem., June 10, 2005; 280(23): 22258 - 22269. [Abstract] [Full Text] [PDF]