Arrestins were originally discovered as negative regulators of G protein-coupled receptor (GPCR) signaling via G proteins. Recent discoveries show that the arrestin-receptor complex initiates signaling through distinct G protein-independent pathways, including those that regulate cell death, survival, and proliferation via MAP kinases. Faulty regulation of GPCR signaling induced by mutations or environmental insults underlies many human diseases. Unfortunately, therapeutic targeting receptor signaling via arrestins, which are natural GPCR regulators, is hampered by lack of selectivity of the non- visual subtypes, both of which interact with dozens of GPCRs. Here we propose to construct and functionally characterize non-visual arrestins with dramatically enhanced specificity for individual GPCRs. Receptor-specific mutants, as well as their enhanced phosphorylation-independent variants, will be tested for their ability to selectively regulate signaling by particuar GPCR subtypes via G proteins, arrestins, and receptor trafficking. Mutants that preferentially interact with particular receptors will be tested for their ability to selectively disrupt receptor coupling to cognate G proteins. Mutants with high preference for D1 and D2 dopamine receptors will be used to determine which receptor subtype plays key role in the development of dyskinesia, the most common devastating side effect of current anti- parkinsonian therapy. Receptor-specific regulation of GPCR signaling has therapeutic potential in multiple disorders associated with congenital or acquired imbalances in cell signaling.

Public Health Relevance

This proposal focuses on the construction and functional characterization of non-visual arrestins with dramatically enhanced specificity for individual G protein-coupled receptors (GPCRs). Receptor-specific mutants will be tested for their ability to selectively regulate signaling by particular GPCR subtypes via G proteins and arrestins, and to determine dopamine receptor subtype that plays key role in dyskinesia, a common side effect of currently used treatment for Parkinson's disease. Receptor-specific regulation of GPCR signaling has therapeutic potential in multiple disorders associated with congenital or acquired imbalances in cell signaling.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM109955-02
Application #
8985683
Study Section
Molecular and Integrative Signal Transduction Study Section (MIST)
Program Officer
Dunsmore, Sarah
Project Start
2015-01-01
Project End
2018-12-31
Budget Start
2016-01-01
Budget End
2016-12-31
Support Year
2
Fiscal Year
2016
Total Cost
$334,515
Indirect Cost
$121,448
Name
Vanderbilt University Medical Center
Department
Pharmacology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
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
Tso, Shih-Chia; Chen, Qiuyan; Vishnivetskiy, Sergey A et al. (2018) Using two-site binding models to analyze microscale thermophoresis data. Anal Biochem 540-541:64-75
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
Zurkovsky, Lilia; Sedaghat, Katayoun; Ahmed, M Rafiuddin et al. (2017) Arrestin-2 and arrestin-3 differentially modulate locomotor responses and sensitization to amphetamine. Neuropharmacology 121:20-29
Chen, Qiuyan; Perry, Nicole A; Vishnivetskiy, Sergey A et al. (2017) Structural basis of arrestin-3 activation and signaling. Nat Commun 8:1427
Indrischek, Henrike; Prohaska, Sonja J; Gurevich, Vsevolod V et al. (2017) Uncovering missing pieces: duplication and deletion history of arrestins in deuterostomes. BMC Evol Biol 17:163
Wanka, Lizzy; Babilon, Stefanie; Burkert, Kerstin et al. (2017) C-terminal motif of human neuropeptide Y4 receptor determines internalization and arrestin recruitment. Cell Signal 29:233-239

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