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

doi:10.1016/S0965-1748(99)00067-3

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 journal › Article › peer-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 language	English
Pages (from-to)	915-930
Number of pages	16
Journal	Insect Biochemistry and Molecular Biology
Volume	29
Issue number	10
DOIs	https://doi.org/10.1016/S0965-1748(99)00067-3
State	Published - 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.

Funding

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.

Funders	Funder number
ZENECA Strategic Research Fund and Zentrum fur Molekulare Neurobiologie

Keywords

Cloning
Ecdysone
Gene expression
Tobacco bud worm
Ultraspiracle

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Insect Science

Access to Document

10.1016/S0965-1748(99)00067-3

Cite this

@article{fcca5f0896e44e4cb3f092b7891d5df2,

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

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.",

keywords = "Cloning, Ecdysone, Gene expression, Tobacco bud worm, Ultraspiracle",

author = "Alberto Martinez and David Scanlon and Barbara Gross and Perara, \{Srini C.\} and Palli, \{Subba R.\} and Greenland, \{Andrew J.\} and John Windass and Olaf Pongs and Peter Broad and Ian Jepson",

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.",

year = "1999",

month = oct,

doi = "10.1016/S0965-1748(99)00067-3",

language = "English",

volume = "29",

pages = "915--930",

number = "10",

}

TY - JOUR

T1 - Transcriptional activation of the cloned Heliothis virescens (Lepidoptera) ecdysone receptor (HvEcR) by Muristerone A

AU - Martinez, Alberto

AU - Scanlon, David

AU - Gross, Barbara

AU - Perara, Srini C.

AU - Palli, Subba R.

AU - Greenland, Andrew J.

AU - Windass, John

AU - Pongs, Olaf

AU - Broad, Peter

AU - Jepson, Ian

N1 - 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.

PY - 1999/10

Y1 - 1999/10

N2 - 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.

AB - 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.

KW - Cloning

KW - Ecdysone

KW - Gene expression

KW - Tobacco bud worm

KW - Ultraspiracle

UR - http://www.scopus.com/inward/record.url?scp=0033215012&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033215012&partnerID=8YFLogxK

U2 - 10.1016/S0965-1748(99)00067-3

DO - 10.1016/S0965-1748(99)00067-3

M3 - Article

C2 - 10528411

AN - SCOPUS:0033215012

SN - 0965-1748

VL - 29

SP - 915

EP - 930

JO - Insect Biochemistry and Molecular Biology

JF - Insect Biochemistry and Molecular Biology

IS - 10

ER -

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

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this