Substitution of synthetic chimpanzee androgen receptor for human androgen receptor in competitive binding and transcriptional activation assays for EDC screening.
The potential effect of receptor-mediated endocrine modulators across species is of increasing concern. In attempts to address these concerns, we are developing androgen and estrogen receptor binding assays using recombinant hormone receptors from a number of species across different vertebrate classes. The United States Environmental Protection Agency (USEPA) Office of Science Coordination and Policy (OSCP) requested that we develop a nonhuman mammalian receptor-binding assay for possible use in their Endocrine Disruptor Screening Program (EDSP). Since the chimpanzee androgen receptor is very similar to that of humans and thus possesses properties which could be exploited in future endocrine studies, we synthesized and expressed this gene in eukaryotic expression plasmids, baculovirus expression vectors and replication deficient adenovirus. In all ligand-binding and transcriptional activation assays tested, the chimpanzee receptor performed essentially identically to the human receptor. This suggests that the chimpanzee gene could substitute for the human gene in endocrine screening assays.
Hartig
P C
PC
Reproductive Toxicology Division, 2525 East Highway 54, MD 72, USEPA, ORD, NHEERL, RTP, NC 27711, United States. hartig.phillip@epa.gov
Cardon
M C
MC
Lambright
C R
CR
Bobseine
K L
KL
Gray
L E
LE
Wilson
V S
VS
D000256:Adenoviridae / Q000235:genetics; D000728:Androgens / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001667:Binding, Competitive; D001681:Biological Assay; D019556:COS Cells; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D052244:Endocrine Disruptors / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D015741:Metribolone / Q000378:metabolism; D002679:Pan troglodytes; D010957:Plasmids; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D015533:Transcriptional Activation; D014161:Transduction, Genetic
C410798:AR protein, human; D000728:Androgens; D052244:Endocrine Disruptors; D011944:Receptors, Androgen; D011994:Recombinant Proteins; D015741:Metribolone