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Dual reporter systems in yeast and mammalian cells for assessing progesterone receptor modulators
Author(s) -
Rudakoff Bärbel,
Undisz Katrin,
Mayer Gerd,
Sobek Lothar,
Kaufmann Günter,
Thiericke Ralf,
Grabley Susanne,
Munder Thomas
Publication year - 1999
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19990401)73:1<126::aid-jcb14>3.0.co;2-e
Subject(s) - reporter gene , saccharomyces cerevisiae , biology , activator (genetics) , receptor , yeast , progesterone receptor , microbiology and biotechnology , promoter , gene , cell culture , gene expression , biochemistry , estrogen receptor , genetics , cancer , breast cancer
In the present study we describe the set‐up of a new one‐hybrid reporter gene assay in Saccharomyces cerevisiae composed of the human progesterone receptor fused to the DNA‐binding domain of the yeast transcriptional activator Gal4. This assay allows the convenient estimation of receptor mediated progestogenic as well as antiprogestogenic actions of compounds. The induction of the β‐galactosidase reporter gene expression correlated well with the progesterone receptor affinity and the concentration of the progestins tested. The results corresponded to those obtained from a reporter gene assay in the cancer cell line CV‐1 and in vitro binding experiments using rabbit uterus cytosol. In both the yeast and CV‐1 cells the activity of antiprogestins was detectable by inhibition of the progestin‐induced reporter gene expression. Secondary reporter genes under the transcriptional control of receptor unrelated promoters have been introduced into yeast and mammalian test strains to distinguish between specific receptor mediated antihormone actions and nonspecific effects on cellular metabolism. J. Cell. Biochem. 73:126–136, 1999. © 1999 Wiley‐Liss, Inc.