Source:http://linkedlifedata.com/resource/pubmed/id/16702214
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
30
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pubmed:dateCreated |
2006-7-24
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pubmed:databankReference | |
pubmed:abstractText |
Incubation of 3T3-L1 preadipocytes with isobutylmethylxanthine (IBMX), dexamethasone, and insulin, alone or in combination, demonstrated that IBMX, which increased cAMP-response element-binding protein (CREB) phosphorylation, was the predominant regulator of Pde3b expression. Real time PCR and immunoblotting indicated that in 3T3-L1 preadipocytes, IBMX-stimulated induction of Pde3b mRNA and protein was markedly inhibited by dominant-negative CREB proteins. By transfecting preadipocytes, differentiating preadipocytes, and HEK293A cells with luciferase reporter vectors containing different fragments of the 5'-flanking region of the Pde3b gene, we identified a distal promoter that contained canonical cis-acting cAMP-response elements (CRE) and a proximal, GC-rich promoter region, which contained atypical CRE. Mutation of the CRE sequences dramatically reduced distal promoter activity; H89 inhibited IBMX-stimulated CREB phosphorylation and proximal and distal promoter activities. Distal promoter activity was stimulated by IBMX and phorbol ester (PMA) in Raw264.7 monocytes, but only by IBMX in 3T3-L1 preadipocytes. Chromatin immunoprecipitation analyses with specific antibodies against CREB, phospho-CREB, and CBP/p300 (CREB-binding protein) showed that these proteins associated with both distal and proximal promoters and that interaction of phospho-CREB, the active form of CREB, with both Pde3b promoter regions was increased in IBMX-treated preadipocytes. These results indicate that CRE in distal and proximal promoter regions and activation of CREB proteins play a crucial role in transcriptional regulation of Pde3b expression during preadipocyte differentiation.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/1-Methyl-3-isobutylxanthine,
http://linkedlifedata.com/resource/pubmed/chemical/3',5'-Cyclic-AMP Phosphodiesterases,
http://linkedlifedata.com/resource/pubmed/chemical/Cyclic AMP Response...,
http://linkedlifedata.com/resource/pubmed/chemical/Cyclic Nucleotide...,
http://linkedlifedata.com/resource/pubmed/chemical/Lipids,
http://linkedlifedata.com/resource/pubmed/chemical/PDE3B protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Pde3b protein, mouse,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphodiesterase Inhibitors
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
28
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pubmed:volume |
281
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
21096-113
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pubmed:dateRevised |
2007-11-15
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pubmed:meshHeading |
pubmed-meshheading:16702214-1-Methyl-3-isobutylxanthine,
pubmed-meshheading:16702214-3',5'-Cyclic-AMP Phosphodiesterases,
pubmed-meshheading:16702214-3T3-L1 Cells,
pubmed-meshheading:16702214-Adipocytes,
pubmed-meshheading:16702214-Animals,
pubmed-meshheading:16702214-Cell Differentiation,
pubmed-meshheading:16702214-Cyclic AMP Response Element-Binding Protein,
pubmed-meshheading:16702214-Cyclic Nucleotide Phosphodiesterases, Type 3,
pubmed-meshheading:16702214-Gene Expression Regulation, Enzymologic,
pubmed-meshheading:16702214-Humans,
pubmed-meshheading:16702214-Lipids,
pubmed-meshheading:16702214-Mice,
pubmed-meshheading:16702214-Models, Genetic,
pubmed-meshheading:16702214-Molecular Sequence Data,
pubmed-meshheading:16702214-Phosphodiesterase Inhibitors
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pubmed:year |
2006
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pubmed:articleTitle |
Importance of cAMP-response element-binding protein in regulation of expression of the murine cyclic nucleotide phosphodiesterase 3B (Pde3b) gene in differentiating 3T3-L1 preadipocytes.
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pubmed:affiliation |
Pulmonary/Critical Care Medicine Branch, NHLBI, National Institutes of Health, Bethesda, Maryland 20892, USA.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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