Jiang, Lily I. and Collins, Julie and Davis, Richard and Lin, Keng-Mean and DeCamp, Dianne and Roach, Tamara and Hsueh, Robert and Rebres, Robert A. and Ross, Elliott M. and Taussig, Ronald and Fraser, Iain and Sternweis, Paul C. (2007) Use of a cAMP BRET Sensor to Characterize a Novel Regulation of cAMP by the Sphingosine 1-Phosphate/G13 Pathway. Journal of Biological Chemistry, 282 (14). pp. 10576-10584. ISSN 0021-9258 http://resolver.caltech.edu/CaltechAUTHORS:JIAjbc07
See Usage Policy.
PDF (Supplementary Data)
See Usage Policy.
Use this Persistent URL to link to this item: http://resolver.caltech.edu/CaltechAUTHORS:JIAjbc07
Regulation of intracellular cyclic adenosine 3',5'-monophosphate (cAMP) is integral in mediating cell growth, cell differentiation, and immune responses in hematopoietic cells. To facilitate studies of cAMP regulation we developed a BRET (bioluminescence resonance energy transfer) sensor for cAMP, CAMYEL (cAMP sensor using YFP-Epac-RLuc), which can quantitatively and rapidly monitor intracellular concentrations of cAMP in vivo. This sensor was used to characterize three distinct pathways for modulation of cAMP synthesis stimulated by presumed Gs-dependent receptors for isoproterenol and prostaglandin E2. Whereas two ligands, uridine 5'-diphosphate and complement C5a, appear to use known mechanisms for augmentation of cAMP via Gq/calcium and Gi, the action of sphingosine 1-phosphate (S1P) is novel. In these cells, S1P, a biologically active lysophospholipid, greatly enhances increases in intracellular cAMP triggered by the ligands for Gs-coupled receptors while having only a minimal effect by itself. The enhancement of cAMP by S1P is resistant to pertussis toxin and independent of intracellular calcium. Studies with RNAi and chemical perturbations demonstrate that the effect of S1P is mediated by the S1P2 receptor and the heterotrimeric G13 protein. Thus in these macrophage cells, all four major classes of G proteins can regulate intracellular cAMP.
|Additional Information:||Copyright © 2007 by the American Society for Biochemistry and Molecular Biology. Received for publication, October 16, 2006 , and in revised form, January 29, 2007. Originally published In Press as doi:10.1074/jbc.M609695200 on February 5, 2007 We thank Jana Hadas for help with protein purification and Drs. William Singer and Scott Gibson for plasmids. This work was supported by National Institutes of Health Grant GM 62114, the Robert A. Welch foundation (to P.C.S.), and the Alfred and Mabel Gilman Chair in Molecular Pharmacology (to P.C.S.). 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 supplemental data and Figs. S1-S7.|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Archive Administrator|
|Deposited On:||22 Aug 2007|
|Last Modified:||26 Dec 2012 09:40|
Repository Staff Only: item control page