Arrestins were discovered as negative regulators of G protein-coupled receptor (GPCR) signaling via G proteins. New data show that the free and receptor-bound arrestins initiate signaling through MAP kinases, which regulate cell death, survival, and proliferation. For example, free and receptor-bound arrestin-3 scaffolds ASK1-MKK4/7-JNK1/2/3 cascades, promoting the activation of JNK family kinases. The two non-visual arrestins interact with ~800 different GPCRs in humans. We propose to identify arrestin elements responsible for receptor specificity, and elucidate the structural basis of the assembly of multi-protein signaling complexes (signalosomes) organized by arrestins. Our new crystal structure of arrestin-3 in the presence of an abundant cytoplasmic molecule IP6 revealed receptor-bound-like (active) conformation of arrestin-3. The ability of arrestin-3 to assume this conformation in the absence of GPCRs likely explains receptor-independent scaffolding activity of arrestin-3. We identified arrestin-3 elements critical for JNK and ERK activation, as well as conformational requirements of distinct branches of arrestin-mediated signaling. We constructed arrestin-3 mutants that bind ASK1, MKK and JNK, but do not promote JNK activation and identified short arrestin-3 peptides that enhance or suppress JNK activity in cells. We will test the potential of signaling-biased arrestins and arrestin-derived molecular tools to facilitate cell death or survival. Molecular tools that specifically increase or block pro-apoptotic signaling have therapeutic potential in disorders associated with excessive cell proliferation (e.g., cancer) or death (e.g., neurodegenerative diseases). Using arrestin-3 mutant specific for dopamine D1 receptor we showed that arrestin-mediated signaling from D1 plays a role in the behavioral sensitization to L-DOPA and development of L-DOPA-induced diskinesia in mouse model of Parkinson?s disease, but other GPCRs also contribute to these phenomena. We believe that signaling-biased arrestins and arrestinbased molecular tools with specific functional capability will help elucidate the intricacies of cellular signaling and yield novel potent therapeutic tools.
We propose to focus on the elucidation of the structural basis of arrestin preference for particular GPCRs, arrestin-dependent activation of pro-apoptotic JNK family kinases, pro-survival ERK1/2 kinases, and upstream activators of these pathways using biochemistry, biophysics, X-ray crystallography, cell- based assays, and experiments in living mice. We propose to test the potential of signaling-biased arrestins to connect to GPCRs of choice pro-survival and pro-apoptotic signaling in cells. Molecular tools that specifically increase or block pro-apoptotic signaling have therapeutic potential in disorders associated with excessive cell proliferation (eg, cancer) or death (eg, neurodegenerative diseases).
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| Tóth, András D; Prokop, Susanne; Gyombolai, Pál et al. (2018) Heterologous phosphorylation-induced formation of a stability lock permits regulation of inactive receptors by ?-arrestins. J Biol Chem 293:876-892 |
| Gurevich, Vsevolod V; Gurevich, Eugenia V (2018) GPCRs and Signal Transducers: Interaction Stoichiometry. Trends Pharmacol Sci 39:672-684 |
| Cleghorn, Whitney M; Bulus, Nada; Kook, Seunghyi et al. (2018) Non-visual arrestins regulate the focal adhesion formation via small GTPases RhoA and Rac1 independently of GPCRs. Cell Signal 42:259-269 |
| Gurevich, Vsevolod V; Gurevich, Eugenia V; Uversky, Vladimir N (2018) Arrestins: structural disorder creates rich functionality. Protein Cell 9:986-1003 |
| Chen, Qiuyan; Iverson, Tina M; Gurevich, Vsevolod V (2018) Structural Basis of Arrestin-Dependent Signal Transduction. Trends Biochem Sci 43:412-423 |
| Zhu, Lu; Rossi, Mario; Cui, Yinghong et al. (2017) Hepatic ?-arrestin 2 is essential for maintaining euglycemia. J Clin Invest 127:2941-2945 |
| Prokop, Susanne; Perry, Nicole A; Vishnivetskiy, Sergey A et al. (2017) Differential manipulation of arrestin-3 binding to basal and agonist-activated G protein-coupled receptors. Cell Signal 36:98-107 |
| Gurevich, Vsevolod V; Gurevich, Eugenia V (2017) Molecular Mechanisms of GPCR Signaling: A Structural Perspective. Int J Mol Sci 18: |
| Vishnivetskiy, Sergey A; Lee, Regina J; Zhou, X Edward et al. (2017) Functional role of the three conserved cysteines in the N domain of visual arrestin-1. J Biol Chem 292:12496-12502 |
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