Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Membranes prime the RapGEF EPAC1 to transduce cAMP signaling

ABSTRACT: EPAC1, a cAMP-activated GEF for Rap GTPases, is a major transducer of cAMP signaling in normal cells and a therapeutic target in cardiac diseases. The recent discovery that cAMP is compartmentalized in membrane-proximal nanodomains challenged the current model of EPAC1 activation in the cytosol. Here, we discover that anionic membranes are a major component of EPAC1 activation. We find that anionic membranes activate EPAC1 in the absence of cAMP, that they increase its affinity for cAMP by 2 orders of magnitude, and that they synergize with cAMP to yield maximal GEF activity. Thus, EPAC1 has four states that differ in their affinity for cAMP and GEF efficiency. In cells, where cytosolic cAMP is low, this implies that EPAC1 must be primed by membranes to bind cAMP. Examination of the inhibitory mechanism of the cardiomyocyte-active chemical CE3F4 in this new framework further reveals that it targets only only membrane- and cAMP-activated EPAC1. Together, our findings reformulate previous concepts of cAMP signaling to include a hitherto overlooked role of membranes through EPAC proteins, with important implications for drug discovery.

INSTRUMENT(S): Synapt MS

ORGANISM(S): Homo Sapiens (human)

DISEASE(S): Cardiovascular System Disease

SUBMITTER: Marie LEY

LAB HEAD: CIANFERANI Sarah

PROVIDER: PXD040227 | Pride | 2023-04-07

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	EPAC_vs_EPAC_AMPc_PML_semiauto_0.5_1_2_5_15min_traite_final.DnX	Other
	checksum.txt	Txt
	cluster_data.csv	Csv
	dataset_EPAC1.zip	Other
	dataset_EPAC1_PML_cAMP.zip	Other

Items per page:

1 - 5 of 7

Similar Datasets

Transcription profiling of mouse bone marrow-derived dendritic cells stimilated with LPS in the presence or absence of cAMP

Project description:genes regualted by LPS or LPS+cAMP stimulation in BMDCs; We used microarrays to identify genes that up-regulated by LPS+cAMP compared with just LPS. Experiment Overall Design: BMDCs were stimilated with LPS (10 ng/ml) in the presence or absence of cAMP (100 microM) for 3h. Specifically up-regualted gene by cMP was identified.

2009-07-05 | E-GEOD-16761 | biostudies-arrayexpress

Dynamics of allosteric regulation of the phospholipase C-gamma isoform

Project description:we used hydrogen deuterium exchange masss spec (HDX-MS) to define the exchange kinetics of PLC-y1 as it interacts with the kinase domain of FGFR1, and liposomes containing PIP2, in order to understand how PLC-y isozymes operate at membranes. We show that the binding of FGFR1k to PLC-y1 increases deuterium incorporation within the interface between the regulatory domains and the catalytic core. Importantly, the addition of liposomes further increase deuterium exchange within the same region.

2022-06-14 | PXD030492 | Pride

Transcription profiling of mouse S49 cells to study the global impact of cAMP and protein kinase A on gene expression

Project description:Although a substantial number of hormones and drugs increase cellular cAMP levels, the global impact of cAMP and its major effector mechanism, protein kinase A (PKA), on gene expression is not known. Here we show that treatment of wild-type S49 lymphoma cells for 24 h with 8-(4-chlorophenylthio)-cAMP (CPT-cAMP), a PKA-selective cAMP analog, alters the expression of ~4500 of ~13,600 unique genes. By contrast, gene expression was unaltered in Kin- S49 cells (that lack PKA) incubated with CPT-cAMP. Changes in mRNA and protein expression of several cell-cycle regulators accompanied cAMP-induced G1-phase cell-cycle arrest of wild-type S49 cells. Within 2 h, CPT-cAMP altered expression of 152 genes that contain evolutionarily conserved cAMP-response elements (CRE) within 5 kb of transcriptional start sites, including the circadian clock gene Per1. Thus, cAMP through its activation of PKA produces extensive transcriptional regulation in eukaryotic cells. These transcriptional networks include a primary group of CRE-containing genes and secondary networks that include the circadian clock.

2007-08-31 | E-GEOD-2413 | biostudies-arrayexpress

Genomic analysis of SRC-2/TIF2 regulated genes in breast cancer cells_

Project description:We have characterized the global program of transcription in SRC-2-depleted MCF-7 breast cancer cells using short-hairpin RNA technology, and in MCF-7 cells exposed to PKA activating agents. In order to identify genes that may be regulated through PKA-induced downregulation of SRC-2, overlapping transcriptional targets in response to the respective treatments were characterized. MCF-7 human breast cancer cells transduced with shRNA targeting SRC-2 (SRC-2 shRNA) or infected with control shRNA vector (Ctr shRNA) were seeded in 9,2 cm petri dishes, 2,0 x106 cells in each dishes, in DMEM supplemented with 5% charcoaled stripped FBS and 10 nM 17b-estradiol. After 48 hours a selection of the Ctr shRNA cells were treated with cAMP analog (8-CPT-cAMP, 150 µM) and cAMP elevating agents (Forskolin (10 µM) and IBMX (50 µM)) The cells were then incubated for another 24 hour in the cell incubator. Cells were harvested by washing them twice with 8 ml PBS (37 oC), before adding 1,5 ml 0,25 % trypzine. The trypzine reaction was stopped after 5 minutes by adding 2,5 ml DMEM to the cells. Trypzine and cell medium was removed by centrifugating the cell suspensions at 1000 rpm for 3 minutes. The cells were then washed with 400 µl PBS, before addition of 350 µl RLT buffer to the cell pellets. The cells were then homogenized by vortexing for 1 minute and then freezed in -80 oC.

2013-05-15 | E-MTAB-1662 | biostudies-arrayexpress

Gene expression induced in HEK293 by treatment with the differentiating agents all-trans retinoic acid (tRA) and N6,2'-O-dibutyryl-cAMP (cAMP)

Project description:This gene array analysis shows that the transcriptome of HEK293, representing low-differentiated human embryonic kidney cells, cultured in presence of tRA and cAMP differentiating agents, is consistent with a genetic program relevant to kidney development Comparison of HEK293 cells grown in standard contitions or in presence of differentiating agents. Cells were seeded in 10 cm dishes, cultured for 3 days either in complete medium or in medium supplemented with tRA and cAMP, harvested, and spun down before RNA isolation. Comparative analysis allowed us to identify genes transcriptionally by tRA/cAMP.

2016-09-06 | E-GEOD-62722 | biostudies-arrayexpress

cAR1 rescues developmental gene expression defects of rasC/rasG double mutants

Project description:It has been shown previously that rasC- cells do not aggregate, predominantly as a result of a defective cAMP relay, although there are also subtle defects in cAMP chemotaxis. There is no appreciable effect on early gene expression in response to starvation. In contrast, rasG-/rasC- cells are totally blocked in aggregation, cAMP relay and cAMP chemotaxis and, in addition, there is no induction of carA and gpaB gene expression, in response to starvation. However, expression of carA from the actin15 promoter in vegetative rasG-/rasC- cells restores the induced expression of carA and gpaB in response to starvation, but has no effect on the defects in aggregation, cAMP chemotaxis and cAMP relay. In order to determine the generality of the effect on early gene expression, micro array analysis was undertaken. These results revealed 121 genes that were significantly (p<0.05) induced in expression 5 fold or more in wild type cells, in response to starvation. Of these genes, the expression of 100 was not induced significantly in rasG-/rasC- cells, but the expression of most of these was restored in rasG-/rasC-/[act15:carA] cells, in response to starvation. These results indicate that the inhibition of early gene expression in rasG-/rasC- cells and its restoration in response to the expression of carA in vegetative cells is a fairly general phenomenon. Thus, signaling through RasG or RasC is required for the induction of expression of many genes that occurs in response to starvation, but the constitutive expression of carA is able to bypass this requirement.

2010-10-31 | E-TABM-1085 | biostudies-arrayexpress

cAMP/PKA signaling balances respiratory activity with mitochondria dependent apoptosis via transcriptional regulation

Project description:This data provides evidence that elevation of cAMP levels has a dramatic effect on the transcriptome of yeast cells, with particular emphasis on mitochondrial function and the promotion of ROS production S. cerevisiae cells lacking the high affinity phosphodiesterase PDE2or both PDE2 and the PKA subunit TPK3 were grown for 24h in rich media in the presence of 4mM exogensously added cAMP. 8 samples.

2010-11-24 | E-GEOD-25541 | biostudies-arrayexpress

Expression data from cAMP-treated WT or IFN-gR1-deficient T cells

Project description:cAMP inhibits TCR signaling, T cell proliferation, cytokine production and T cell function. We used microarrays to detail the global programme of gene expression in TCR-activated WT or IFN-gR1-deficient CD4+ T cells by db-cAMP. CD4+ T cells were purified from spleens of WT or IFN-gR1-deficient mice by autoMACS, and activated with anti-CD3 plus anti-CD28 for 15 h. db-cAMP or vehicle control (med) was added into cultures for the last 3 h.

2012-07-23 | E-GEOD-39592 | biostudies-arrayexpress

Genomics of signalling crosstalk of estrogen receptor alpha in breast cancer cells

Project description:The estrogen receptor a (ERa) is a ligand-regulated transcription factor. However, a wide variety of other extracellular signals can activate ERa in the absence of estrogen. The impact of these alternate modes of activation on gene expression profiles has not been characterized. We show that estrogen, growth factors and cAMP elicit surprisingly distinct ERa-dependent transcriptional responses in human MCF7 breast cancer cells. In response to growth factors and cAMP, ERa primarily activates and represses genes, respectively. The combined treatments with the antiestrogen tamoxifen and cAMP or growth factors regulate yet other sets of genes. In many cases, tamoxifen is perverted to an agonist, potentially mimicking what is happening in certain tamoxifen-resistant breast tumors and emphasizing the importance of the cellular signaling environment. Using a computational analysis, we predicted that a Hox protein might be involved in mediating such combinatorial effects, and then confirmed it experimentally. Although both tamoxifen and cAMP block the proliferation of MCF7 cells, their combined application stimulates it, and this can be blocked with a dominant-negative Hox mutant. Thus, the activating signal dictates both target gene selection and regulation by ERa, and this has consequences on global gene expression patterns that may be relevant to understanding the progression of ERa-dependent carcinomas. Keywords: treatment response 10 treatment conditions (3 biological replicates for each, totalling 30 individual samples), of which 3 were untreated controls. Each replicate was hybridized to a separate chip, totalling 27 cDNA slides representing 9 unique conditions.

2008-05-22 | E-GEOD-10466 | biostudies-arrayexpress

Interaction of cGMP-dependent protein kinase and HAP1

Project description:Here, we use cAMP-based chemical proteomics approach, employing competitive concentrations of free cyclic nucleotides to isolate PKA and PKG and their secondary interactors.

2015-02-09 | PXD001434 | Pride