Chimeric receptors as a tool for luminescent measurement of biological activities of steroid hormones.
Some applications of chimeric cellular models are presented to study the biological activities of steroid hormones. We have used several chimeric constructs encoding the DNA binding domain of Gal4 yeast protein fused to the hormone binding domain of various steroid receptors (MR, PR, GR and ER). Interactions of these chimeric receptors with a 17-mer DNA sequence, specific for Gal-4, control expression of the firefly luciferase as a reporter gene. Stable transfected cell lines expressing the firefly luciferase under the control of different steroids were established and an efficient and easy sub-cloning was allowed with the help of an imaging system using a single-photon-counting camera. In the cell lines obtained, the bioluminescent response can be easily measured and thus used to measure specific biological activities of steroid agonists or antagonists. We observed that the responses are effector-concentration-dependent and their biological activities will be compared to those of native receptors.
Jausons-Loffreda
N
N
INSERM Unité 58, Montpellier, France.
Balaguer
P
P
Roux
S
S
Fuentes
M
M
Pons
M
M
Nicolas
J C
JC
Gelmini
S
S
Pazzagli
M
M
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D001517:Coleoptera / Q000201:enzymology; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis; D005656:Fungal Proteins / Q000096:biosynthesis / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000096:biosynthesis*; D008163:Luminescent Measurements; D010957:Plasmids; D011987:Receptors, Steroid / Q000096:biosynthesis / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D013256:Steroids / Q000494:pharmacology*; D014157:Transcription Factors; D014162:Transfection
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C052130:GAL4 protein, S cerevisiae; D011987:Receptors, Steroid; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D013256:Steroids; D014157:Transcription Factors; D004247:DNA; D008156:Luciferases