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Article: Induction of chinook salmon growth hormone promoter activity by the adenosine 3',5'-monophosphate (cAMP)-dependent pathway involves two cAMP- response elements with the CGTCA motif and the pituitary-specific transcription factor Pit-1

TitleInduction of chinook salmon growth hormone promoter activity by the adenosine 3',5'-monophosphate (cAMP)-dependent pathway involves two cAMP- response elements with the CGTCA motif and the pituitary-specific transcription factor Pit-1
Authors
Issue Date1996
PublisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.org
Citation
Endocrinology, 1996, v. 137 n. 5, p. 1775-1784 How to Cite?
AbstractIn this study, the functional role of two cAMP-response elements (CRE) in the promoter of the chinook salmon GH gene and their interactions with the transcription factor Pit-1 in regulating GH gene expression were examined. A chimeric construct of the chloramphenicol acetyltransferase (CAT) reporter gene with the CRE-containing GH promoter (pGH.CAT) was transiently transfected into primary cultures of rainbow trout pituitary cells. The expression of CAT activity was stimulated by an adenylate cyclase activator forskolin as well as a membrane-permeant cAMP analog 8-bromo-cAMP. Furthermore, these stimulatory responses were inhibited by a protein kinase A inhibitor H89, suggesting that these CREs are functionally coupled to the adenylate cyclase-cAMP-protein kinase A cascade. This hypothesis is supported by parallel studies using GH 4ZR 7 cells, a rat pituitary cell line stably transfected with dopamine D2 receptors. In this cell line, D2 receptor activation is known to inhibit adenylate cyclase activity and cAMP synthesis. Stimulation with a nonselective dopamine agonist, apomorphine, or a D2- specific agonist, Ly171555, suppressed the expression of pGH.CAT in GH 4ZR 7 cells, and this inhibition was blocked by simultaneous treatment with forskolin. These results indicate that inhibition of the cAMP-dependent pathway reduces the basal promoter activity of the CRE-containing pGH-.CAT. The functionality of these CREs was further confirmed by deletion analysis and site-specific mutagenesis. In trout pituitary cells, the cAMP inducibility of pGH.CAT was inhibited after deleting the CRE-containing sequence from the GH promoter. When the CRE-containing sequence was cloned into a CAT construct with a vital thymidine kinase promoter, a significant elevation of cAMP inducibility was observed. This stimulatory response, however, was abolished by mutating the core sequence, CGTCA, in these CREs, suggesting that these cis-acting elements confer cAMP inducibility to the salmon GH gene. The interactions between CREs and the transcription factor Pit-1 in mediating GH gene expression were also examined. In HeLa cells, a human cervical cancer cell line deficient in Pit-1, both basal and cAMP- induced expression of pGH.CAT were apparent only with the cotransfection of a Pit-1 expression vector. These results taken together indicate that the two CREs in the chinook salmon GH gene are functionally associated with the cAMP- dependent pathway and that their promoter activity is dependent on the presence of Pit-1.
Persistent Identifierhttp://hdl.handle.net/10722/49428
ISSN
2015 Impact Factor: 4.159
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DC FieldValueLanguage
dc.contributor.authorWong, AOLen_HK
dc.contributor.authorLe Drean, Yen_HK
dc.contributor.authorLiu, Den_HK
dc.contributor.authorZhi Zhou Huen_HK
dc.contributor.authorShao Jun Duen_HK
dc.contributor.authorHew, CLen_HK
dc.date.accessioned2008-06-12T06:42:26Z-
dc.date.available2008-06-12T06:42:26Z-
dc.date.issued1996en_HK
dc.identifier.citationEndocrinology, 1996, v. 137 n. 5, p. 1775-1784en_HK
dc.identifier.issn0013-7227en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49428-
dc.description.abstractIn this study, the functional role of two cAMP-response elements (CRE) in the promoter of the chinook salmon GH gene and their interactions with the transcription factor Pit-1 in regulating GH gene expression were examined. A chimeric construct of the chloramphenicol acetyltransferase (CAT) reporter gene with the CRE-containing GH promoter (pGH.CAT) was transiently transfected into primary cultures of rainbow trout pituitary cells. The expression of CAT activity was stimulated by an adenylate cyclase activator forskolin as well as a membrane-permeant cAMP analog 8-bromo-cAMP. Furthermore, these stimulatory responses were inhibited by a protein kinase A inhibitor H89, suggesting that these CREs are functionally coupled to the adenylate cyclase-cAMP-protein kinase A cascade. This hypothesis is supported by parallel studies using GH 4ZR 7 cells, a rat pituitary cell line stably transfected with dopamine D2 receptors. In this cell line, D2 receptor activation is known to inhibit adenylate cyclase activity and cAMP synthesis. Stimulation with a nonselective dopamine agonist, apomorphine, or a D2- specific agonist, Ly171555, suppressed the expression of pGH.CAT in GH 4ZR 7 cells, and this inhibition was blocked by simultaneous treatment with forskolin. These results indicate that inhibition of the cAMP-dependent pathway reduces the basal promoter activity of the CRE-containing pGH-.CAT. The functionality of these CREs was further confirmed by deletion analysis and site-specific mutagenesis. In trout pituitary cells, the cAMP inducibility of pGH.CAT was inhibited after deleting the CRE-containing sequence from the GH promoter. When the CRE-containing sequence was cloned into a CAT construct with a vital thymidine kinase promoter, a significant elevation of cAMP inducibility was observed. This stimulatory response, however, was abolished by mutating the core sequence, CGTCA, in these CREs, suggesting that these cis-acting elements confer cAMP inducibility to the salmon GH gene. The interactions between CREs and the transcription factor Pit-1 in mediating GH gene expression were also examined. In HeLa cells, a human cervical cancer cell line deficient in Pit-1, both basal and cAMP- induced expression of pGH.CAT were apparent only with the cotransfection of a Pit-1 expression vector. These results taken together indicate that the two CREs in the chinook salmon GH gene are functionally associated with the cAMP- dependent pathway and that their promoter activity is dependent on the presence of Pit-1.en_HK
dc.format.extent418 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.orgen_HK
dc.relation.ispartofEndocrinologyen_HK
dc.rightsEndocrinology. Copyright © The Endocrine Society.en_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshCyclic AMP - pharmacologyen_HK
dc.subject.meshDNA - chemistry - metabolismen_HK
dc.subject.meshDNA-Binding Proteins - metabolismen_HK
dc.subject.meshGrowth Hormone - geneticsen_HK
dc.subject.meshPromoter Regions (Genetics)en_HK
dc.titleInduction of chinook salmon growth hormone promoter activity by the adenosine 3',5'-monophosphate (cAMP)-dependent pathway involves two cAMP- response elements with the CGTCA motif and the pituitary-specific transcription factor Pit-1en_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0013-7227&volume=137&issue=5&spage=1775&epage=1784&date=1996&atitle=Induction+of+chinook+salmon+growth+hormone+promoter+activity+by+the+adenosine+3%27,5%27-monophosphate+(cAMP)-dependent+pathway+involves+two+cAMP-response+elements+with+the+CGTCA+motif+and+the+pituitary-specific+transcription+factor+Pit-1en_HK
dc.identifier.emailWong, AOL: olwong@hkucc.hku.hken_HK
dc.identifier.authorityWong, AOL=rp00806en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1210/en.137.5.1775en_HK
dc.identifier.pmid8612514en_HK
dc.identifier.scopuseid_2-s2.0-0029863663en_HK
dc.identifier.hkuros11885-
dc.identifier.hkuros26606-
dc.identifier.volume137en_HK
dc.identifier.issue5en_HK
dc.identifier.spage1775en_HK
dc.identifier.epage1784en_HK
dc.identifier.isiWOS:A1996UG63600037-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridWong, AOL=7403147570en_HK
dc.identifier.scopusauthoridLe Drean, Y=6603392595en_HK
dc.identifier.scopusauthoridLiu, D=36706226300en_HK
dc.identifier.scopusauthoridZhi Zhou Hu=8152519400en_HK
dc.identifier.scopusauthoridShao Jun Du=7409732054en_HK
dc.identifier.scopusauthoridHew, CL=7007113098en_HK

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