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

Bovine steroid 21-hydroxylase: regulation of biosynthesis.

John ME, Okamura T, Dee A, Adler B, John MC, White PC, Simpson ER, Waterman MR
Biochemistry. 1986 25 (10): 2846-53

PMID: 2424492 · DOI:10.1021/bi00358a016

A recombinant cDNA clone, PBC21-1, specific for bovine steroid 21-hydroxylase cytochrome P-450 (P-450C21) was identified in a bovine adrenocortical cDNA library, and this identity was confirmed by nucleotide sequencing which revealed significant amino acid homology (77%) with human P-450C21 cDNA. The pBC21-1 insert is 1.7 kilobases in length and includes a 1128 base pair region that encodes the C-terminal 376 amino acids of bovine P-450C21 as well as 535 base pairs of 3'-untranslated sequence. A novel feature of this insert is a 20 base pair intervening sequence near the 5' end, apparently the result of an aberrant splicing event. Northern blot analysis reveals that bovine P-450C21 is encoded by two transcripts, 2.3 and 2.0 kilobases in length which are detected in adrenal cortical RNA. Bovine liver, heart, kidney, and corpus luteum do not contain detectable P-450C21 transcripts. Regulation of P-450C21 gene expression by adrenocorticotropin was investigated with pBC21-1 and bovine adrenocortical cells in primary, monolayer culture. Treatment with ACTH or analogues of cAMP increases the steady-state levels of P-450C21 RNA in such cell cultures. In vitro transcription run-on assays suggest that this increase is, at least in part, due to the enhanced transcriptional activity of the P-450C21 gene.

MeSH Terms (22)

Adrenal Cortex Amino Acid Sequence Animals Base Sequence Cattle Cells, Cultured Cloning, Molecular Cytochrome P-450 Enzyme System DNA DNA Restriction Enzymes Humans Kinetics Mice Nucleic Acid Hybridization Organ Specificity RNA Sequence Homology, Nucleic Acid Species Specificity Steroid 21-Hydroxylase Steroid Hydroxylases Swine Transcription, Genetic

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