Evidence map›Paper›PMID 25913012›Full record

ArticleBiochimica et biophysica acta2015

Phosphorylation of ezrin on Thr567 is required for the synergistic activation of cell spreading by EPAC1 and protein kinase A in HEK293T cells.

Euan Parnell, Andreas Koschinski, Manuela Zaccolo, Ryan T Cameron, George S Baillie, Gemma L Baillie, Alison Porter, Stuart P McElroy, Stephen J Yarwood

Open access · hybridAbstract read
In one paragraph

Article in Biochimica et biophysica acta, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.

0numbers the graph read from it
0cells of the map it votes in
10citing papers in PubMed
1.2field-weighted citation impact, top 21% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

10 citing papers in PubMed, 18 citations in OpenAlex.

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  10. Ezrin Orchestrates Signal Transduction in Airway Cells.Reviews of physiology, biochemistry and pharmacology
    Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors at 2 institutions in 1 country.

Euan ParnellInstitute of Molecular, Cellular and Systems Biology, College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Andreas KoschinskiDepartment of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK.
Manuela ZaccoloDepartment of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK.
Ryan T CameronInstitute of Cardiovascular and Medical Sciences, College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
George S BaillieInstitute of Cardiovascular and Medical Sciences, College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Gemma L BaillieEuropean Screening Centre, BioCity Scotland, Newhouse ML1 5UH, UK.
Alison PorterEuropean Screening Centre, BioCity Scotland, Newhouse ML1 5UH, UK.
Stuart P McElroyEuropean Screening Centre, BioCity Scotland, Newhouse ML1 5UH, UK.
Stephen J YarwoodInstitute of Molecular, Cellular and Systems Biology, College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK. Electronic address: stephen.yarwood@glasgow.ac.uk.
University of Glasgow · GBUniversity of Oxford · GB

Funding

Biotechnology and Biological Sciences Research Council BB/D015324/1Biotechnology and Biological Sciences Research Council BB/F016735/1British Heart Foundation PG/10/26/28303British Heart Foundation PG/15/15/31316British Heart Foundation RG/12/3/29423Medical Research Council MR/J007412/1
6 · The paper itself

Abstract

Recent studies have demonstrated that the actin binding protein, ezrin, and the cAMP-sensor, EPAC1, cooperate to induce cell spreading in response to elevations in intracellular cAMP. To investigate the mechanisms underlying these effects we generated a model of EPAC1-dependent cell spreading based on the stable transfection of EPAC1 into HEK293T (HEK293T-EPAC1) cells. We found that direct activation of EPAC1 with the EPAC-selective analogue, 8-pCPT-2'-O-Me-cAMP (007), promoted cell spreading in these cells. In addition, co-activation of EPAC1 and PKA, with a combination of the adenylate cyclase activator, forskolin, and the cAMP phosphodiesterase inhibitor, rolipram, was found to synergistically enhance cell spreading, in association with cortical actin bundling and mobilisation of ezrin to the plasma membrane. PKA activation was also associated with phosphorylation of ezrin on Thr567, as detected by an electrophoretic band mobility shift during SDS-PAGE. Inhibition of PKA activity blocked ezrin phosphorylation and reduced the cell spreading response to cAMP elevation to levels induced by EPAC1-activation alone. Transfection of HEK293T-EPAC1 cells with inhibitory ezrin mutants lacking the key PKA phosphorylation site, ezrin-Thr567Ala, or the ability to associate with actin, ezrin-Arg579Ala, promoted cell arborisation and blocked the ability of EPAC1 and PKA to further promote cell spreading. The PKA phospho-mimetic mutants of ezrin, ezrin-Thr567Asp had no effect on EPAC1-driven cell spreading. Our results indicate that association of ezrin with the actin cytoskeleton and phosphorylation on Thr567 are required, but not sufficient, for PKA and EPAC1 to synergistically promote cell spreading following elevations in intracellular cAMP.

Indexed as

Cell MovementAnimalsCell MembraneChlorocebus aethiopsCOS CellsCyclic AMPCyclic AMP-Dependent Protein KinasesCytoskeletal ProteinsCytoskeletonEzrinGenes, DominantGuanine Nucleotide Exchange FactorsHEK293 CellsHumansHuman Umbilical Vein Endothelial CellsMicrofilament ProteinsCyclic AMPCyclic AMP-Dependent Protein KinasesCytoskeletal ProteinsEzrinGuanine Nucleotide Exchange FactorsMicrofilament ProteinsPhosphothreonineProtein Kinase InhibitorsRAPGEF3 protein, humanCell morphologyCyclic AMPCytoskeletonEPAC1

Identifiers

PMID25913012
PMCPMC4547084
OpenAlexW2044576721

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.