PMID- 7982913
OWN - NLM
STAT- MEDLINE
DCOM- 19941230
LR  - 20210212
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 269
IP  - 48
DP  - 1994 Dec 2
TI  - The 90-kDa heat shock protein is essential for Ah receptor signaling in a yeast 
      expression system.
PG  - 30109-12
AB  - In an effort to provide a more powerful system to study the Ah receptor (AHR) 
      signaling pathway, we expressed the AHR, its dimerization partner ARNT, and a 
      beta-galactosidase (lacZ) reporter gene, driven by two dioxin-responsive 
      enhancers, in the yeast Saccharomyces cerevisiae. In this system, the agonists 
      beta-naphthoflavone and alpha-naphthoflavone induced transcription of the lacZ 
      gene, with EC50 values of 7.9 x 10(-8) and 3.0 x 10(-7) M, respectively, while 
      the nonagonist dexamethasone was without effect. As a first application of this 
      system, we examined the relationship between the 90-kDa heat shock protein 
      (hsp90) and AHR function. To accomplish this in a manner that was independent of 
      the ARNT protein, we constructed a chimeric receptor in which the DNA binding and 
      primary dimerization domains of the AHR were swapped with analogous domains from 
      the LexA protein. Coexpression of this AHR-LexA chimera and a lacZ reporter gene 
      driven by eight LexA operator sites in a yeast strain with regulatable levels of 
      hsp90, yielded pharmacology that closely mirrored that of the 
      AHR/ARNT/dioxin-responsive enhancer system described above, but only when hsp90 
      levels were held near their wild type levels. When hsp90 levels were reduced to 
      approximately 5% of normal, AHR signaling in response to agonist was completely 
      blocked despite normal cell growth. These results provide the first genetic 
      evidence for the role of hsp90 in AHR signaling and provide the basis for a 
      powerful new system in which to study this pathway.
FAU - Carver, L A
AU  - Carver LA
AD  - Department of Molecular Pharmacology and Biological Chemistry, Northwestern 
      University Medical School, Chicago, Illinois 60611.
FAU - Jackiw, V
AU  - Jackiw V
FAU - Bradfield, C A
AU  - Bradfield CA
LA  - eng
GR  - ES05703/ES/NIEHS NIH HHS/United States
GR  - T32-ES07124/ES/NIEHS NIH HHS/United States
GR  - T32-GM08061/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Bacterial Proteins)
RN  - 0 (Benzoflavones)
RN  - 0 (DNA Primers)
RN  - 0 (HSP90 Heat-Shock Proteins)
RN  - 0 (LexA protein, Bacteria)
RN  - 0 (Receptors, Aryl Hydrocarbon)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 604-59-1 (alpha-naphthoflavone)
RN  - 6051-87-2 (beta-Naphthoflavone)
RN  - 7S5I7G3JQL (Dexamethasone)
RN  - EC 3.2.1.23 (beta-Galactosidase)
RN  - EC 3.4.21.- (Serine Endopeptidases)
SB  - IM
MH  - Bacterial Proteins/biosynthesis
MH  - Base Sequence
MH  - Benzoflavones/pharmacology
MH  - DNA Primers
MH  - Dexamethasone/pharmacology
MH  - Gene Expression/drug effects
MH  - Genes, Bacterial/drug effects
MH  - HSP90 Heat-Shock Proteins/biosynthesis/*metabolism
MH  - Humans
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction
MH  - Receptors, Aryl Hydrocarbon/agonists/biosynthesis/*physiology
MH  - Recombinant Fusion Proteins/biosynthesis
MH  - Restriction Mapping
MH  - Saccharomyces cerevisiae/growth & development/*physiology
MH  - *Serine Endopeptidases
MH  - *Signal Transduction
MH  - Temperature
MH  - *Transcription, Genetic/drug effects
MH  - beta-Galactosidase/biosynthesis
MH  - beta-Naphthoflavone
EDAT- 1994/12/02 00:00
MHDA- 1994/12/02 00:01
CRDT- 1994/12/02 00:00
PHST- 1994/12/02 00:00 [pubmed]
PHST- 1994/12/02 00:01 [medline]
PHST- 1994/12/02 00:00 [entrez]
AID - S0021-9258(18)43782-9 [pii]
PST - ppublish
SO  - J Biol Chem. 1994 Dec 2;269(48):30109-12.