A CRE-like sequence that binds CREB and contributes to cAMP-dependent regulation of the proximal promoter of the human aromatase P450 (CYP19) gene.
The major physiological regulator of human aromatase P450 gene expression in the ovary is follicle stimulating hormone (FSH), which acts by increasing intracellular cAMP levels. This study describes the identification of an element in the aromatase proximal promoter that is critical for the full transcriptional response of this promoter to cAMP. The cAMP-responsive element (CRE)-like sequence (CLS) was originally identified by its sequence similarity to a palindromic CRE, from which it differs by the insertion of a single cytosine. Mutation of the CLS in the context of 278 bp of 5'-flanking DNA resulted in the loss of cAMP-induced reporter gene expression in transfected ovarian luteal cells. A cell line survey EMSA revealed that CLS binding factors are ubiquitously distributed, although the migration pattern of CLS-nuclear protein complexes varied among different nuclear extracts. An extended half-site for binding members of the basic-leucine zipper class of transcription factors was found to be responsible for ovarian luteal cell nuclear protein binding and cAMP-dependent transcriptional transactivation. Competition and supershift EMSAs revealed that the CLS-nuclear protein complexes that regulate cAMP-induced transcription were indistinguishable from homodimeric CREB bound to the CRE oligonucleotide, yet the interaction with the CLS was of lower affinity.
Michael
M D
MD
Cecil H. and Ida Green Center for Reproductive Biology Sciences, The University of Texas Southwestern Medical Center, Dallas 75235-9051, USA.
Michael
L F
LF
Simpson
E R
ER
D000818:Animals; D001141:Aromatase / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D003338:Corpus Luteum / Q000166:cytology / Q000201:enzymology; D000242:Cyclic AMP / Q000378:metabolism* / Q000502:physiology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D005260:Female; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic / Q000187:drug effects
D017362:Cyclic AMP Response Element-Binding Protein; D009687:Nuclear Proteins; D009841:Oligonucleotides; D005576:Colforsin; D000242:Cyclic AMP; D008156:Luciferases; D001141:Aromatase