# Targeted Engineering of Designer Arrestins to Regulate Cell Signaling

> **NIH NIH R35** · VANDERBILT UNIVERSITY · 2020 · $564,977

## Abstract

PROJECT SUMMARY
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.

## Key facts

- **NIH application ID:** 9914303
- **Project number:** 5R35GM122491-04
- **Recipient organization:** VANDERBILT UNIVERSITY
- **Principal Investigator:** VSEVOLOD V. GUREVICH
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $564,977
- **Award type:** 5
- **Project period:** 2017-05-01 → 2022-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9914303

## Citation

> US National Institutes of Health, RePORTER application 9914303, Targeted Engineering of Designer Arrestins to Regulate Cell Signaling (5R35GM122491-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9914303. Licensed CC0.

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