Michael Waterman
Faculty Member
Last active: 2/12/2015

Transcriptional regulation of the bovine CYP17 (P-450(17)alpha) gene. Identification of two cAMP regulatory regions lacking the consensus cAMP-responsive element (CRE).

Lund J, Ahlgren R, Wu DH, Kagimoto M, Simpson ER, Waterman MR
J Biol Chem. 1990 265 (6): 3304-12

PMID: 1689300

Regions within the 5'-flanking sequence of the bovine CYP17 (P-450(17)alpha) gene which are required for cAMP-dependent regulation of transcription have been localized by transient transfection of chimeric reporter gene constructs into mouse adrenal tumor Y1 cells. Two sequences have been found which individually confer cAMP responsiveness to reporter genes; they are located at -243/-225 and -80/-40 base pairs (bp). Obvious sequence homology between these two regions is not apparent. Gel shift competition analysis indicates that nuclear protein(s) binding to the -243/-225-bp region can be competed for by the addition of a double-stranded oligonucleotide containing a consensus cAMP-responsive element (CRE) from the human chorionic gonadotropin alpha gene, whereas addition of this CRE does not abolish protein-DNA complexes formed with fragments containing the -80/-40-bp sequence. Gel shift and Southwestern analysis indicate that the -243/-225-bp region of the P-450(17)alpha gene and the CRE both bind a 47-kDa protein and that the CRE binds additional proteins (43 and 68 kDa) not apparently recognized by the -243/-225-bp sequence. Thus cAMP-dependent regulation of the bovine P-450(17)alpha gene appears to involve two independent cis-regulatory regions, neither of which contains a consensus CRE. Based on protein binding analysis, one of these regions (that including -80/-40 bp) is distinct from the consensus CRE while the other (that containing -243/-225 bp) may be related to the consensus CRE.

MeSH Terms (23)

Adrenal Gland Neoplasms Animals Cattle Chimera Chorionic Gonadotropin Chromosome Deletion Cyclic AMP Response Element-Binding Protein DNA-Binding Proteins Gene Amplification Gene Expression Regulation, Enzymologic Genes Genes, Regulator Humans Immunoblotting Mice Nuclear Proteins Oligonucleotide Probes Promoter Regions, Genetic RNA Steroid 17-alpha-Hydroxylase Steroid Hydroxylases Transcription, Genetic Transfection

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