Exchange protein directly activated by cyclic AMP-1-regulated recruitment of CCAAT/enhancer-binding proteins to the suppressor of cytokine signaling-3 promoter

Sands, W. A., Woolson, H. D., Yarwood, S. J. and Palmer, T. M. (2012) Exchange protein directly activated by cyclic AMP-1-regulated recruitment of CCAAT/enhancer-binding proteins to the suppressor of cytokine signaling-3 promoter. Methods in Molecular Biology, 809, pp. 201-214. (doi: 10.1007/978-1-61779-376-9_14)

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Publisher's URL: http://dx.doi.org/10.1007/978-1-61779-376-9_14

Abstract

The ability of prototypical second messenger cyclic AMP (cAMP) to positively control transcription of the somatostatin gene was pivotal to the original identification of the transcription factor cAMP response element-binding protein. However, it is now clear that alternative intracellular cAMP sensors, of which the exchange protein directly activated by cAMP (Epac) proteins have been studied most intensively, also initiate transcription of key genes in response to cAMP elevation. For example, we have demonstrated in vascular endothelial cells that activation of Epac1 is necessary for cAMP-mobilizing agents to trigger the induction of the gene-encoding suppressor of cytokine signaling-3 (SOCS-3), a potent inhibitor of interleukin (IL)-6 signaling. This is achieved through the recruitment of CCAAT/enhancer-binding protein (C/EBP) transcription factors to the SOCS-3 promoter. Here, we describe in detail how to identify and measure cAMP-mediated recruitment of a specific C/EBP isoform to a candidate regulator region of the SOCS-3 promoter in vascular endothelial cells in vitro. We also describe the RNA interference strategies with which we identified a role for Epac1 and SOCS-3 in being responsible for mediating the inhibitory effect of cAMP elevation on IL-6 signaling.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Yarwood, Dr Stephen and Palmer, Dr Timothy and Sands, Dr William
Authors: Sands, W. A., Woolson, H. D., Yarwood, S. J., and Palmer, T. M.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Methods in Molecular Biology
Publisher:Humana Press (Springer Imprint)
ISSN:1064-3745
ISSN (Online):1940-6029

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
498581EPAC1 and ERK-dependent activation of C/EBP transcription factors: a new cyclic AMP-activated anti-inflammatory gene expression module in vascular endothelial cellsTimothy PalmerBritish Heart Foundation (BHF)PG/08/125/26415RI CARDIOVASCULAR & MEDICAL SCIENCES