title: N-formyl peptide receptors internalize but do not recycle in the absence of arrestins.
abstract: Arrestins mediate phosphorylation-dependent desensitization, internalization, and initiation of signaling cascades for the majority of G protein-coupled receptors (GPCRs). Many GPCRs undergo agonist-mediated internalization through arrestin-dependent mechanisms, wherein arrestin serves as an adapter between the receptor and endocytic proteins. To understand the role of arrestins in N-formyl peptide receptor (FPR) trafficking, we stably expressed the FPR in a mouse embryonic fibroblast cell line (MEF) that lacked endogenous arrestin 2 and arrestin 3 (arrestin-deficient). We compared FPR internalization and recycling kinetics in these cells to congenic wild type MEF cell lines. Internalization of the FPR was not altered in the absence of arrestins. Since the FPR remains associated with arrestins following internalization, we investigated whether the rate of FPR recycling was altered in arrestin-deficient cells. While the FPR was able to recycle in the wild type cells, receptor recycling was largely absent in the arrestin double knockout cells. Reconstitution of the arrestin-deficient line with either arrestin 2 or arrestin 3 restored receptor recycling. Confocal fluorescence microscopy studies demonstrated that in arrestin-deficient cells the FPR may become trapped in the perinuclear recycling compartment. These observations indicate that, although the FPR can internalize in the absence of arrestins, recycling of internalized receptors to the cell surface is prevented. Our results suggest a novel role for arrestins in the post-endocytic trafficking of GPCRs.
authors.affiliation: Department of Cell Biology & Physiology, The University of New Mexico Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA.
authors.forename: Charlotte M
authors.initials: CM
authors.lastname: Vines
authors.forename: Chetana M
authors.initials: CM
authors.lastname: Revankar
authors.forename: Diane C
authors.initials: DC
authors.lastname: Maestas
authors.forename: Leah L
authors.initials: LL
authors.lastname: LaRusch
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Cimino
authors.forename: Trudy A
authors.initials: TA
authors.lastname: Kohout
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Lefkowitz
authors.forename: Eric R
authors.initials: ER
authors.lastname: Prossnitz
mesh_terms: D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D004705:Endocytosis; D007700:Kinetics; D051379:Mice; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D010750:Phosphoproteins / Q000235:genetics; D021381:Protein Transport; D044042:Receptors, Formyl Peptide / Q000378:metabolism*; D014162:Transfection
chemical_list: D019390:Arrestins; D010750:Phosphoproteins; D044042:Receptors, Formyl Peptide; C082333:arrestin3