Pons, MónicaIzquierdo, IsmaelAndreu-Carbó, MireiaGarrido, GeorginaPlanagumà, JesúsMuriel, OliviaPozo, Miguel Ángel delIsabel Geli, M.Aragay, Anna M.2018-06-152018-06-152017Pons M, Izquierdo I, Andreu-Carbo M, Garrido G, Planaguma J, Muriel O et al. Phosphorylation of filamin A regulates chemokine receptor CCR2 recycling. J Cell Sci. 2017 Jan 15;130(2):490-501. DOI: 10.1242/jcs.193821. Epub 2016 Dec 10021-9533http://hdl.handle.net/10230/34914Proper endosomal trafficking of ligand-activated G-protein-coupled receptors (GPCRs) is essential to spatiotemporally tune their physiological responses. For the monocyte chemoattractant receptor 2 (CCR2B; one of two isoforms encoded by CCR2), endocytic recycling is important to sustain monocyte migration, whereas filamin A (FLNa) is essential for CCL2-induced monocyte migration. Here, we analyze the role of FLNa in the trafficking of CCR2B along the endocytic pathway. In FLNa-knockdown cells, activated CCR2B accumulated in enlarged EEA-1-positive endosomes, which exhibited slow movement and fast fluorescence recovery, suggesting an imbalance between receptor entry and exit rates. Utilizing super-resolution microscopy, we observed that FLNa-GFP, CCR2B and β2-adrenergic receptor (β2AR) were present in actin-enriched endosomal microdomains. Depletion of FLNa decreased CCR2B association with these microdomains and concomitantly delayed CCR2B endosomal traffic, without apparently affecting the number of microdomains. Interestingly, CCR2B and β2AR signaling induced phosphorylation of FLNa at residue S2152, and this phosphorylation event was contributes to sustain receptor recycling. Thus, our data strongly suggest that CCR2B and β2AR signals to FLNa to stimulate its endocytosis and recycling to the plasma membrane.application/pdfengPublished originally by Cold Spring Harbor Laboratory Press at 10.1242/jcs.193821. Beginning six months from the full-issue publication date, articles are distributed under the Creative Commons Attribution-Non-Commercial 4.0 International License (CC-BY-NC), as described at http://creativecommons.org/licenses/by-nc/4.0/. This license permits non-commercial use, including reproduction, adaptation, and distribution of the article provided the original author and source are credited.info:eu-repo/semantics/articlehttp://dx.doi.org/10.1242/jcs.193821G-protein-coupled receptorsCcr2Filamin ARecyclinginfo:eu-repo/semantics/openAccess