A cAMP-responsive element regulates expression of the mouse steroid 11 beta-hydroxylase gene.
In Y1 mouse adrenocortical tumor cells, expression of steroid 11 beta-hydroxylase (11 beta-OHase) is stimulated by cAMP following a delay of 4-6 h. Our results demonstrate that a cAMP-responsive element (CRE) within the 11 beta-OHase promoter region is a major determinant of this induction. The 5'-flanking sequences from the mouse 11 beta-OHase gene were placed in front of a human growth hormone reporter gene and transfected into Y1 cells. Treatment of transfected cells with 8-bromo-cAMP increased expression directed by the 11 beta-OHase 5'-flanking region by 3.8-fold. In 5'-deletion analyses, 123 base pairs of 5'-flanking sequences were sufficient for cAMP induction, whereas cAMP treatment did not affect expression of a plasmid with only 40 base pairs of 5'-flanking sequence. Within these 123 base pairs, a region from -56 to -49 matched 7 of 8 bases comprising the consensus sequence for the CRE. 11 beta-OHase 5'-flanking sequences from -65 to -42, including the CRE-like sequence, conferred cAMP inducibility to promoters from the thymidine kinase and chorionic gonadotropin alpha-subunit genes. DNase I footprinting and Southwestern blotting analyses demonstrated that the protein which interacted with the CRE in the 11 beta-OHase promoter region was similar to the CRE-binding protein associated with other cAMP-regulated genes. Together, these results suggest that an interaction between the 11 beta-OHase CRE and CRE-binding protein mediates cAMP induction of the 11 beta-OHase gene.
Rice
D A
DA
Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710.
Aitken
L D
LD
Vandenbark
G R
GR
Mouw
A R
AR
Franklin
A
A
Schimmer
B P
BP
Parker
K L
KL
D000306:Adrenal Cortex Neoplasms / Q000235:genetics; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification; D000242:Cyclic AMP / Q000378:metabolism / Q000494:pharmacology*; D002373:Cyclic AMP Receptor Protein; D003850:Deoxyribonuclease I; D005786:Gene Expression Regulation / Q000187:drug effects*; D005796:Genes; D006367:HeLa Cells; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000235:genetics / Q000302:isolation & purification; D009710:Nucleotide Mapping; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016760:Proto-Oncogene Proteins c-fos; D013252:Steroid 11-beta-Hydroxylase / Q000235:genetics*; D013250:Steroid Hydroxylases / Q000235:genetics*; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D016760:Proto-Oncogene Proteins c-fos; D000242:Cyclic AMP; D013250:Steroid Hydroxylases; D013252:Steroid 11-beta-Hydroxylase; D003850:Deoxyribonuclease I