GPCR signaling is essential for a wide variety of physiological processes. GPCR signaling can also play an active role in the pathogenesis of multiple diseases. GPCRs are normally tightly regulated so that signals are of the appropriate magnitude and duration and any perturbations in these regulatory processes can be deleterious. Typically, GPCR signaling is tightly regulated by G protein-coupled receptor kinases (GRKs) and ?-arrestins. ?-arrestins bind to GRK-phosphorylated GPCRs at the plasma membrane to terminate signaling by promoting G protein uncoupling and receptor endocytosis. The molecular mechanisms by which GRKs and ?- arrestins regulate GPCR signaling remain poorly understood. The goal of this proposal is to elucidate the molecular mechanisms by which GRKs and ?-arrestins govern GPCR signaling. Recently, we described a novel function for ?-arrestins, as endosomal sorting molecules, whereby they function on endosomes to mediate sorting of GPCRs from endosomes to lysosomes, leading to receptor degradation and long-term attenuation of signaling. However, mechanistic insight into this new function is lacking. ?-arrestin-1 mediates endosomal sorting of the chemokine receptor CXCR4, which also requires the action of the E3 ubiquitin ligase AIP4 and the ESCRT (endosomal sorting complex required for transport) machinery. How ?-arrestin-1 functionally integrates with AIP4 and ESCRTs to mediate CXCR4 endosomal sorting remains to be determined. Based on our strong preliminary data we hypothesize that ?-arrestin-1 regulates the ESCRT machinery to control CXCR4 endosomal trafficking and signaling. We propose the following two specific aims:
Aim 1 : To determine the molecular mechanisms by which ?-arrestin-1 integrates with AIP4 and ESCRTs to control CXCR4 endosomal sorting.
Aim 2 : To determine the molecular mechanisms by which ESCRTs regulate CXCR4 signaling and function. We will utilize a comprehensive series of state-of-the-art biochemical, molecular and cellular biology and imaging approaches to complete these aims. We anticipate that new knowledge gained from this proposal will lead to the identification of new and innovative approaches to manipulate GPCR signaling to prevent and treat a variety of diseases.

Public Health Relevance

The goals of this proposal are to characterize and elucidate the molecular mechanisms by which beta arrestins govern endosomal sorting of G protein-coupled receptors. GPCR signaling has to be of the appropriate magnitude and duration as any perturbations in GPCR signaling may be pathogenic. Although beta arrestins play an important role in regulating GPCR signaling, their role in endosomal sorting remains poorly understood. Understanding how beta arrestins govern GPCR endosomal sorting may lead to the identification of new and innovative therapeutic targets to treat and prevent diseases in which GPCR signaling is involved.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Molecular Neuropharmacology and Signaling Study Section (MNPS)
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Ainsztein, Alexandra M
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Loyola University Chicago
Schools of Medicine
United States
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Nevins, Amanda M; Marchese, Adriano (2018) Detecting Cell Surface Expression of the G Protein-Coupled Receptor CXCR4. Methods Mol Biol 1722:151-164
English, Elizabeth J; Mahn, Sarah A; Marchese, Adriano (2018) Endocytosis is required for CXC chemokine receptor type 4 (CXCR4)-mediated Akt activation and antiapoptotic signaling. J Biol Chem 293:11470-11480
Kennedy, Justine E; Marchese, Adriano (2015) Regulation of GPCR Trafficking by Ubiquitin. Prog Mol Biol Transl Sci 132:15-38
Verma, Rita; Marchese, Adriano (2015) The endosomal sorting complex required for transport pathway mediates chemokine receptor CXCR4-promoted lysosomal degradation of the mammalian target of rapamycin antagonist DEPTOR. J Biol Chem 290:6810-24
Holleman, Justine; Marchese, Adriano (2014) The ubiquitin ligase deltex-3l regulates endosomal sorting of the G protein-coupled receptor CXCR4. Mol Biol Cell 25:1892-904
Marchese, Adriano (2014) Endocytic trafficking of chemokine receptors. Curr Opin Cell Biol 27:72-7