Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-2
pubmed:dateCreated
2001-7-30
pubmed:abstractText
Transient transfection of mouse gonadotrope-derived (alphaT3-1) cells with a 2297 bp human GnRHR promoter-luciferase construct (p2300-LucF) showed a dose- and time-dependent increase in the human gonodotropin-releasing hormone receptor (GnRHR) promoter activity after forskolin treatment. An average of 4.8-fold increase in promoter activity was observed after 12 h of 10 microM forskolin treatment. This effect was mimicked by administration of cholera toxin, cAMP analog or pituitary adenylate cyclase activating polypeptide 38 (PACAP). A specific adenylate cyclase (AC) inhibitor (ACI) or protein kinase A (PKA) inhibitor (PKAI) pretreatment reversed the forskolin- and PACAP-induced increase in the human GnRHR promoter activity. These results not only confirm the stimulatory effect of Cyclic adenosine monophosphate (cAMP) in human GnRHR promoter activation, but also suggest that hormones or neurotransmitters that activate adenylate cyclase in pituitary gonadotropes may increase the expression of human GnRHR gene in transcriptional level. Progressive 5' deletion assays identified a 412 bp fragment (-577 to 167) in the human GnRHR 5'-flanking region that is essential in maintaining the basal responsiveness to cAMP. Mutagenesis coupled with functional studies have identified two putative AP-1/CREB binding sites, namely hGR-AP/CRE-1 and hGR-AP/CRE-2 that participated in mediating the cAMP-stimulatory effect. Mutation of the putative hGR-AP/CRE-1 and hGR-CRE-2 resulted in a 38 and 32% decrease in the forskolin-induced stimulation. However, mutation of both binding sites did not completely abolish the cAMP-stimulatory effect, suggesting that multiple transcription factor binding sites were involved in full response in cAMP stimulation. The binding of CREB to these motifs was confirmed by gel mobility shift assay and antibody supershift assay.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
http://linkedlifedata.com/resource/pubmed/chemical/8-Bromo Cyclic Adenosine..., http://linkedlifedata.com/resource/pubmed/chemical/ADCYAP1 protein, human, http://linkedlifedata.com/resource/pubmed/chemical/Adcyap1 protein, mouse, http://linkedlifedata.com/resource/pubmed/chemical/Adenylate Cyclase, http://linkedlifedata.com/resource/pubmed/chemical/Cholera Toxin, http://linkedlifedata.com/resource/pubmed/chemical/Cyclic AMP, http://linkedlifedata.com/resource/pubmed/chemical/Cyclic AMP-Dependent Protein Kinases, http://linkedlifedata.com/resource/pubmed/chemical/DNA, http://linkedlifedata.com/resource/pubmed/chemical/Forskolin, http://linkedlifedata.com/resource/pubmed/chemical/Neuropeptides, http://linkedlifedata.com/resource/pubmed/chemical/Nuclear Proteins, http://linkedlifedata.com/resource/pubmed/chemical/Pituitary Adenylate..., http://linkedlifedata.com/resource/pubmed/chemical/RNA, Messenger, http://linkedlifedata.com/resource/pubmed/chemical/Receptors, LHRH
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0303-7207
pubmed:author
pubmed:issnType
Print
pubmed:day
5
pubmed:volume
181
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
15-26
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:11476937-8-Bromo Cyclic Adenosine Monophosphate, pubmed-meshheading:11476937-Adenylate Cyclase, pubmed-meshheading:11476937-Animals, pubmed-meshheading:11476937-Cell Line, pubmed-meshheading:11476937-Cholera Toxin, pubmed-meshheading:11476937-Cyclic AMP, pubmed-meshheading:11476937-Cyclic AMP-Dependent Protein Kinases, pubmed-meshheading:11476937-DNA, pubmed-meshheading:11476937-Forskolin, pubmed-meshheading:11476937-Genes, Reporter, pubmed-meshheading:11476937-Humans, pubmed-meshheading:11476937-Mice, pubmed-meshheading:11476937-Mutagenesis, Site-Directed, pubmed-meshheading:11476937-Neuropeptides, pubmed-meshheading:11476937-Nuclear Proteins, pubmed-meshheading:11476937-Pituitary Adenylate Cyclase-Activating Polypeptide, pubmed-meshheading:11476937-Pituitary Gland, pubmed-meshheading:11476937-Promoter Regions, Genetic, pubmed-meshheading:11476937-RNA, Messenger, pubmed-meshheading:11476937-Receptors, LHRH, pubmed-meshheading:11476937-Response Elements, pubmed-meshheading:11476937-Sequence Deletion, pubmed-meshheading:11476937-Signal Transduction, pubmed-meshheading:11476937-Transcription, Genetic, pubmed-meshheading:11476937-Up-Regulation
pubmed:year
2001
pubmed:articleTitle
Human gonadotropin-releasing hormone receptor gene transcription: up-regulation by 3',5'-cyclic adenosine monophosphate/protein kinase A pathway.
pubmed:affiliation
Department of Obstetrics and Gynaecology, The University of British Columbia, B.C. Women's Hospital, 2H30-4490 Oak Street, BC, V6H 3V5, Vancouver, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't