HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 281, Issue 30, 21096-21113, July 28, 2006

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 281/30/21096    most recent M601307200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Liu, H. Articles by Manganiello, V. C. Search for Related Content PubMed PubMed Citation Articles by Liu, H. Articles by Manganiello, V. C. Social Bookmarking                      What's this?

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^*

Hanguan Liu¹, Jing Rong Tang¹, Young Hun Choi, Maria Napolitano, Steven Hockman, Masato Taira, Eva Degerman², and Vincent C. Manganiello³

From the Pulmonary/Critical Care Medicine Branch, NHLBI, National Institutes of Health, Bethesda, Maryland 20892 and Section for Molecular Signaling, Department of Cell and Molecular Biology, University of Lund, S-22100 Lund, Sweden

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.

Received for publication, February 10, 2006 , and in revised form, May 15, 2006.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF547434 [GenBank] , AY159890 [GenBank] , AF547435 [GenBank] , and AAN52086 [GenBank] .

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains additional text and Fig. S1.

¹ Both authors contributed equally to this work.

² Supported in part by the Swedish Medical Research Council Project 3362.

³ To whom correspondence should be addressed: Pulmonary-Critical Care Medicine Branch, NHLBI, National Institutes of Health, Bldg. 10, Rm. 5N307, 10 Center Dr., Bethesda, MD 20892-1434. Tel.: 301-496-1770; Fax: 301-402-1610; E-mail: manganiv{at}nhlbi.nih.gov.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Sasseville, N. Cote, C. Vigneault, C. Guillemette, and F. J. Richard 3'5'-Cyclic Adenosine Monophosphate-Dependent Up-Regulation of Phosphodiesterase Type 3A in Porcine Cumulus Cells Endocrinology, April 1, 2007; 148(4): 1858 - 1867. [Abstract] [Full Text] [PDF]