PMID- 11435614
OWN - NLM
STAT- MEDLINE
DCOM- 20011025
LR  - 20151119
IS  - 0888-8809 (Print)
IS  - 0888-8809 (Linking)
VI  - 15
IP  - 7
DP  - 2001 Jul
TI  - Reconstruction of ligand-dependent transactivation of Choristoneura fumiferana 
      ecdysone receptor in yeast.
PG  - 1140-53
AB  - Ecdysteroids play an important role in regulating development and reproduction in 
      insects. Interaction of 20-hydroxyecdysone (20E) with ecdysone receptor (EcR) as 
      a heterodimer with ultraspiracle (USP) protein triggers the activation of 
      20E-responsive genes. In this paper we describe a ligand-mediated transactivation 
      system in yeast using the spruce budworm Choristoneura fumiferana ecdysone 
      receptor (CfEcR). Coexpression of C. fumiferana USP (CfUSP) with CfEcR in yeast 
      led to constitutive transcription of the reporter gene. However, deletion of the 
      A/B domain of CfUSP abolished constitutive activity observed for the CfUSP:CfEcR 
      complex. Replacement of USP with its mammalian homolog retinoid X receptors 
      (RXRs) abolished the constitutive activity of the heterodimer but it did not 
      restore EcR ligand-mediated transactivation. These data suggest that USP and its 
      A/B domain play a role in the constitutive function of CfEcR:USP in yeast. A 
      ligand-mediated transactivation was observed when GRIP1, a mouse coactivator 
      gene, was added to EcR:RXR or EcR:DeltaA/BUSP complexes. Deletion of the A/B 
      domain of EcR in the context of DeltaA/BEcR:RXR:GRIP1 or 
      DeltaA/BEcR:DeltaA/BUSP:GRIP1 dramatically improved the ligand-dependent 
      transactivation. This is the first example of highly efficient ligand-dependent 
      transactivation of insect EcR in yeast. Analysis of transactivation activity of 
      different ecdysteroidal compounds showed that the yeast system remarkably mimics 
      the response observed in insect tissue culture cells and whole insect systems. 
      The results open the way to develop assays that can be used to screen novel 
      species-specific ecdysone agonist/antagonist insecticides.
FAU - Tran, H T
AU  - Tran HT
AD  - LifeSensors, Inc., Malvern, Pennsylvania 19355, USA.
FAU - Askari, H B
AU  - Askari HB
FAU - Shaaban, S
AU  - Shaaban S
FAU - Price, L
AU  - Price L
FAU - Palli, S R
AU  - Palli SR
FAU - Dhadialla, T S
AU  - Dhadialla TS
FAU - Carlson, G R
AU  - Carlson GR
FAU - Butt, T R
AU  - Butt TR
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Mol Endocrinol
JT  - Molecular endocrinology (Baltimore, Md.)
JID - 8801431
RN  - 0 (Insect Proteins)
RN  - 0 (Ncoa2 protein, mouse)
RN  - 0 (Nuclear Receptor Coactivator 2)
RN  - 0 (Receptors, Retinoic Acid)
RN  - 0 (Receptors, Steroid)
RN  - 0 (Retinoid X Receptors)
RN  - 0 (Transcription Factors)
RN  - 0 (ecdysone receptor)
RN  - 10028-17-8 (Tritium)
RN  - 3604-87-3 (Ecdysone)
RN  - 5289-74-7 (Ecdysterone)
RN  - 84986BG3NG (ponasterone A)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - DNA/metabolism
MH  - Drosophila melanogaster/metabolism
MH  - Ecdysone/agonists/pharmacology
MH  - Ecdysterone/*analogs & derivatives/metabolism/pharmacology
MH  - Escherichia coli/genetics
MH  - Gene Expression
MH  - Insect Proteins/genetics/metabolism
MH  - Larva/drug effects
MH  - Lepidoptera/*genetics
MH  - Mice
MH  - Nuclear Receptor Coactivator 2
MH  - Plasmids/genetics
MH  - Receptors, Retinoic Acid/genetics
MH  - Receptors, Steroid/*genetics
MH  - Retinoid X Receptors
MH  - Saccharomyces cerevisiae/*genetics
MH  - Spodoptera/drug effects
MH  - Transcription Factors/genetics
MH  - *Transcriptional Activation
MH  - Transfection
MH  - Tritium
EDAT- 2001/07/04 10:00
MHDA- 2001/10/26 10:01
CRDT- 2001/07/04 10:00
PHST- 2001/07/04 10:00 [pubmed]
PHST- 2001/10/26 10:01 [medline]
PHST- 2001/07/04 10:00 [entrez]
AID - 10.1210/mend.15.7.0660 [doi]
PST - ppublish
SO  - Mol Endocrinol. 2001 Jul;15(7):1140-53. doi: 10.1210/mend.15.7.0660.