Document Detail


Composite glucocorticoid regulation at a functionally defined negative glucocorticoid response element of the human corticotropin-releasing hormone gene.
MedLine Citation:
PMID:  10517666     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Glucocorticoid-dependent negative feedback of the hypothalamic-pituitary-adrenal axis is mediated in part through direct inhibition of hypothalamic CRH gene transcription. In the present study, we sought to further localize and characterize glucocorticoid receptor (GR) and AP-1 interactions at a functionally defined negative glucocorticoid response element (nGRE) of the CRH promoter. Transient transfection studies in mouse corticotroph AtT-20 cells demonstrated that internal deletion of the nGRE (-278 to -249 nucleotides) within the context of 1 kb of the intact CRH promoter resulted in decreased 8-BrcAMP stimulation and glucocorticoid-dependent repression of CRH promoter activity. The nGRE conferred transcriptional activation by both cAMP and overexpressed c-jun or c-fos AP-1 nucleoproteins as well as specific glucocorticoid-dependent repression to a heterologous promoter. A similar profile of regulation was observed for the composite GRE derived from mouse proliferin promoter. The CRH nGRE was clearly distinct from the consensus cAMP response element (CRE) at -224 nucleotides, which increased basal activity and cAMP responsiveness of a heterologous promoter but did not confer glucocorticoid-dependent repression. High-affinity binding sites for both GR and AP-1 nucleoproteins were identified at adjacent elements within the nGRE. Mutations that disrupted either GR or AP-1 binding activity were associated with loss of glucocorticoid-dependent repression. These results are consistent with a composite mechanism of glucocorticoid-dependent repression involving direct DNA binding of GR and AP-1 nucleoproteins at discrete adjacent sites within the CRH promoter.
Authors:
S P Malkoski; R I Dorin
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Molecular endocrinology (Baltimore, Md.)     Volume:  13     ISSN:  0888-8809     ISO Abbreviation:  Mol. Endocrinol.     Publication Date:  1999 Oct 
Date Detail:
Created Date:  1999-12-22     Completed Date:  1999-12-22     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8801431     Medline TA:  Mol Endocrinol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1629-44     Citation Subset:  IM    
Affiliation:
Department of Medicine, Albuquerque Veterans Administration Medical Center, New Mexico 87108, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Binding Sites
Cell Line
Conserved Sequence
Corticotropin-Releasing Hormone / genetics*,  metabolism
Cyclic AMP / metabolism
Cyclic AMP Response Element-Binding Protein / genetics,  metabolism
Genes, fos
Genes, jun
Glucocorticoids / metabolism*
Humans
Luciferases / genetics,  metabolism
Mice
Mutation
Pituitary Gland, Anterior / cytology,  metabolism
Promoter Regions, Genetic
Receptors, Glucocorticoid / genetics,  metabolism
Recombinant Proteins / genetics,  metabolism
Regulatory Sequences, Nucleic Acid
Response Elements / physiology*
Sequence Deletion
Transcription Factor AP-1 / metabolism
Transcription, Genetic
Chemical
Reg. No./Substance:
0/Cyclic AMP Response Element-Binding Protein; 0/Glucocorticoids; 0/Receptors, Glucocorticoid; 0/Recombinant Proteins; 0/Transcription Factor AP-1; 60-92-4/Cyclic AMP; 9015-71-8/Corticotropin-Releasing Hormone; EC 1.13.12.-/Luciferases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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