Expression of three negative regulators of CYP17 gene transcription in adrenocortical cells.
The expression of negative regulators of CYP17 gene expression: DAX-1, COUP-TF and N-CoR was investigated in bovine adrenocortical cells in primary culture. The cells were incubated for 6 hours in a defined medium, containing ACTH (10 nM) or forskolin (25 microM). Total RNA was isolated, the concentration of CYP17 gene transcript was determined by hybridization analysis, and polyA RNA was reverse-transcribed to obtain cDNA. Fragments of CYP17, DAX-1, COUP-TF, N-CoR and beta-actin cDNA were amplified by PCR and the amplified cDNAs were analyzed by agarose gel electrophoresis followed by densitometry. It was found that DAX-1 and COUP-TF mRNA levels were unaffected by treatment with ACTH or forskolin. On the contrary, both compounds enhanced the level of mRNA encoding N-CoR. Under the same conditions, ACTH or forskolin substantially stimulated CYP 17 mRNA accumulation. Our results suggest that in the adrenocortical cells in culture, the expression of DAX1 and COUP-TF is constitutive, whereas N-CoR expression might be inducible.
Lehmann
T
T
Department of Biochemistry and Molecular Biology, University of Medical Sciences, Poznan, Poland.
Biernacka-Lukanty
J
J
Trzeciak
W H
WH
D000302:Adrenal Cortex / Q000166:cytology / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology; D000818:Animals; D051836:COUP Transcription Factors; D002417:Cattle; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D057137:DAX-1 Orphan Nuclear Receptor; D004268:DNA-Binding Proteins / Q000235:genetics; D004338:Drug Combinations; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D056971:Nuclear Receptor Co-Repressor 1; D012333:RNA, Messenger / Q000378:metabolism; D018168:Receptors, Retinoic Acid / Q000235:genetics; D011987:Receptors, Steroid; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology*
D051836:COUP Transcription Factors; D057137:DAX-1 Orphan Nuclear Receptor; D004268:DNA-Binding Proteins; D004338:Drug Combinations; D009687:Nuclear Proteins; D056971:Nuclear Receptor Co-Repressor 1; D012333:RNA, Messenger; D018168:Receptors, Retinoic Acid; D011987:Receptors, Steroid; D012097:Repressor Proteins; D014157:Transcription Factors; D005576:Colforsin; D000324:Adrenocorticotropic Hormone; D013254:Steroid 17-alpha-Hydroxylase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases