Adrenocortical-specific transgene expression directed by steroid hydroxylase gene promoters.
The 5'-flanking regions of genes for three mouse adrenal steroid hydroxylases were analyzed for their ability to direct adrenal cortex-specific beta-galactosidase (beta-gal) reporter expression both in cell culture and transgenic mice. The 5'-flanking regions chosen were from the genes for steroid 21-hydroxylase (21-OHase), expressed throughout the adrenal cortex and mediating both glucocorticoid and mineralocorticoid synthesis, and aldosterone synthetase (AS) and steroid 11 beta-hydroxylase (11 beta-OHase), which catalyze respectively the terminal steps of mineralocorticoid synthesis in the zona glomerulosa and glucocorticoid synthesis in the zona fasciculata/reticularis. While 5.0 kb of 11 beta-OHase gene 5'-flanking region and 5.4 kb of the AS gene 5'-flanking region mediated respectively moderate and low levels of beta-gal reporter expression in Y1 adrenocortical tumor cells, neither of these 5'-flanking regions was able to direct reporter expression to the appropriate adrenocortical zone of transgenic mice. This suggests that additional regulatory elements, lying outside these 5'-flanking regions, are required for 11 beta-OHase and AS gene expression in the intact mouse. In contrast, 6.4 kb of the mouse 21-OHase A gene 5' flanking region was able to direct specific beta-galactosidase reporter expression, in both Y1 cells and transgenic mice, indicating that elements directing adrenal cortex-specific gene expression in vivo are located not more than 6.4 kb 5' of the 21-OHase gene transcription start site.
Morley
S D
SD
Centre for Genome Research, University of Edinburgh, UK.
Viard
I
I
Parker
K L
KL
Mullins
J J
JJ
D000302:Adrenal Cortex / Q000201:enzymology*; D000818:Animals; D019405:Cytochrome P-450 CYP11B2 / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D013252:Steroid 11-beta-Hydroxylase / Q000235:genetics; D013255:Steroid 21-Hydroxylase / Q000235:genetics; D013250:Steroid Hydroxylases / Q000235:genetics*; D014162:Transfection; D019076:Transgenes; D001616:beta-Galactosidase / Q000235:genetics
D013250:Steroid Hydroxylases; D013255:Steroid 21-Hydroxylase; D019405:Cytochrome P-450 CYP11B2; D013252:Steroid 11-beta-Hydroxylase; D001616:beta-Galactosidase