Adaptor protein-2 interaction with arrestin regulates GPCR recycling and apoptosis.
G protein-coupled receptors (GPCRs) are integral to cellular function in nearly all physiologic and many pathologic processes. GPCR signaling represents an intricate balance between receptor activation, inactivation (desensitization, internalization and degradation) and resensitization (recycling and de novo synthesis). Complex formation between phosphorylated GPCRs, arrestins and an ever-increasing number of effector molecules is known to regulate cellular function. Previous studies have demonstrated that, although N-formyl peptide receptor (FPR) internalization occurs in the absence of arrestins, FPR recycling is arrestin-dependent. Furthermore, FPR stimulation in the absence of arrestins leads to receptor accumulation in perinuclear endosomes and apoptosis. In this study, we show that the interaction of GPCR-bound arrestin with adaptor protein-2 (AP-2) is a critical anti-apoptotic event. In addition, AP-2 associates with the receptor-arrestin complex in perinuclear endosomes and is required for proper post-endocytic GPCR trafficking. Finally, we observed that depletion of endogenous AP-2 results in the initiation of apoptosis upon stimulation of multiple GPCRs, including P2Y purinergic receptors and CXCR2, but not CXCR4. We propose a model in which the abnormal accumulation of internalized GPCR-arrestin complexes in recycling endosomes, resulting from defective arrestin-AP-2 interactions, leads to the specific initiation of aberrant signaling pathways and apoptosis.
Wagener
Brant M
BM
Department of Cell Biology and Physiology and UNM Cancer Research and Treatment Center, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA.
Marjon
Nicole A
NA
Revankar
Chetana M
CM
Prossnitz
Eric R
ER
D033962:Adaptor Protein Complex 2 / Q000235:genetics / Q000378:metabolism*; D017209:Apoptosis / Q000502:physiology*; D019390:Arrestins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D011992:Endosomes / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D021381:Protein Transport; D044042:Receptors, Formyl Peptide / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000096:biosynthesis* / Q000378:metabolism; D014162:Transfection; D020298:U937 Cells
D033962:Adaptor Protein Complex 2; D019390:Arrestins; D044042:Receptors, Formyl Peptide; D043562:Receptors, G-Protein-Coupled; D049452:Green Fluorescent Proteins