Nucleic Acids Research, Vol 26, Issue 15 3577-3583, Copyright © 1998 by Oxford University Press
CA Miller 3rd, MA Martinat and LE Hyman
The pBEVY (bi-directional expression vectors for yeast) plasmids were
designed with constitutive and galactose-induced bi-directional promoters
to direct the expression of multiple proteins in Saccharomyces cerevisiae .
Using human estrogen receptor as a test gene, relatively balanced
expression levels from each side of a bi- directional promoter were
observed. Expression of a functional heterodimeric transcription factor
composed of human aryl hydrocarbon receptor (Ahr) and aryl hydrocarbon
receptor nuclear translocator (Arnt) proteins was accomplished using a
single pBEVY plasmid. Previous studies suggest that inhibitory cross-talk
between the estrogen receptor and the Ahr/Arnt complex may occur and that
Hsp90-Ahr complex formation is important for Ahr-mediated signal
transduction. Evidence for functional interaction among these proteins was
investigated using pBEVY plasmids in a yeast system. No inhibitory
cross-talk was observed in signaling assays performed with yeast that
co-expressed Ahr, Arnt and estrogen receptor. In contrast,
Ahr/Arnt-mediated signal transduction was reduced by 80% in a
temperature-sensitive Hsp90 strain grown under non-permissive conditions.
We conclude that pBEVY plasmids facilitate the examination of multiple
protein interactions in yeast model systems.
ARTICLES
Assessment of aryl hydrocarbon receptor complex interactions using pBEVY plasmids: expressionvectors with bi-directional promoters for use in Saccharomyces cerevisiae
Environmental Health Sciences Department and Tulane-Xavier Center for Bioenvironmental Research,Tulane University School of Public Health and Tropical Medicine, 1430 Tulane Avenue, New Orleans,LA 70112, USA.
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