Transcriptional activation of the cloned Heliothis virescens (Lepidoptera) ecdysone receptor (HvEcR) by Muristerone A

Alberto Martinez, David Scanlon, Barbara Gross, Srini C. Perara, Subba R. Palli, Andrew J. Greenland, John Windass, Olaf Pongs, Peter Broad, Ian Jepson

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

Ecdysteroids play an important role during insect development. We report here the isolation and characterisation of an Ecdysone receptor (EcR) homologue from Heliothis virescens (HvEcR) and present evidence supporting the HvEcR active role as an active component of the native insect receptor. Alignment of the deduced amino acid sequence of HvEcR with those of EcRs from other species confirmed its membership of this family and showed that it is closely related to the B1 isoform of Drosophila melanogaster. Northern blot analysis showed that two transcripts (6.0 and 6.5 kb) were recognised by a probe spanning the DNA and ligand binding domains of the HvEcR. Genomic Southern blots showed that the HvEcR is encoded by a single copy gene. Two lines of evidence towards the functional activity of the HvEcR are presented. In vitro transcribed and translated HvEcR showed specific binding to hsp27 and pall response elements in the presence of CfUSP. Stable expression of HvEcR in 293 cells induced reporter gene activity in the presence of muristerone A in a dose dependant manner while dexamethasone failed to activate.

Original languageEnglish
Pages (from-to)915-930
Number of pages16
JournalInsect Biochemistry and Molecular Biology
Volume29
Issue number10
DOIs
StatePublished - Oct 1999

Bibliographical note

Funding Information:
We would like to thank the ZENECA Strategic Research Fund and Zentrum fur Molekulare Neurobiologie, for support in carrying out this work. Elena Puente is thanked for her kind gift of the random primed larval H. virescens library. Dr. M. Needham is thanked for the gift of the mammalian reporter plasmid. Protein sequence alignments were produced with the help of Dr A. Evans.

Keywords

  • Cloning
  • Ecdysone
  • Gene expression
  • Tobacco bud worm
  • Ultraspiracle

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Insect Science

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