Arrestins were first described 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. In particular, both free and receptor- associated arrestin-3 scaffolds ASK1-MKK-JNK cascade, promoting JNK activation. Here we propose to elucidate the structural basis of arrestin-dependent activation of pro-apoptotic JNK family kinases and their activators MKK4/7 using biochemical and biophysical methods. The molecular mechanisms of the assembly of multi-protein signaling complexes (signalosomes) organized by arrestin-3 will be established, and arrestin-3 residues critical for JNK activation will be identified. Based on ths info, arrestin-3 mutants that bind ASK1, MKK and JNK, but do not promote JNK activation will be constructed. The potential of arrestin mutants with dramatically reduced ability to activate JNKs, several of which we already have, to protect cells against insults and prolong their survival will be tested. We have also constructed arrestin-3 mutants that activate JNKs more efficiently than parental wild type arrestin-3. We established the paradigm where arrestin-3-dependent JNK activation plays key role in cell survival. We will test the ability of hyperactive arrestin-3 mutants to facilitate cell death. We showed that inactive mutants tie up JNKs and upstream kinases in unproductive complexes, thereby acting in dominant- negative manner. We will test the potential of these mutants to protect cells and prolong their survival. Molecular toos 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).

Public Health Relevance

This proposal focuses on the elucidation of the structural basis of arrestin-dependent activation of pro- apoptotic JNK family kinases and their activators MKK4/7 using biochemical and biophysical methods. The potential of arrestin mutants with dramatically reduced ability to activate JNKs, that were constructed based on this info, to protect cells against insults and prolong their survival will be tested, as well as the ability of the mutats that activate JNKs more efficiently than wild type arrestins to facilitate cell death will be also tested. 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).

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM077561-05A1
Application #
8295479
Study Section
Biophysics of Neural Systems Study Section (BPNS)
Program Officer
Dunsmore, Sarah
Project Start
2006-04-01
Project End
2016-03-31
Budget Start
2012-04-10
Budget End
2013-03-31
Support Year
5
Fiscal Year
2012
Total Cost
$306,047
Indirect Cost
$101,949
Name
Vanderbilt University Medical Center
Department
Pharmacology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Chen, Qiuyan; Perry, Nicole A; Vishnivetskiy, Sergey A et al. (2017) Structural basis of arrestin-3 activation and signaling. Nat Commun 8:1427
Zhu, Lu; AlmaƧa, Joana; Dadi, Prasanna K et al. (2017) ?-arrestin-2 is an essential regulator of pancreatic ?-cell function under physiological and pathophysiological conditions. Nat Commun 8:14295
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
Gurevich, Eugenia V; Gainetdinov, Raul R; Gurevich, Vsevolod V (2016) G protein-coupled receptor kinases as regulators of dopamine receptor functions. Pharmacol Res 111:1-16
Zhan, Xuanzhi; Stoy, Henriette; Kaoud, Tamer S et al. (2016) Peptide mini-scaffold facilitates JNK3 activation in cells. Sci Rep 6:21025
Hu, Jianxin; Stern, Matthew; Gimenez, Luis E et al. (2016) A G Protein-biased Designer G Protein-coupled Receptor Useful for Studying the Physiological Relevance of Gq/11-dependent Signaling Pathways. J Biol Chem 291:7809-20
Zhan, Xuanzhi; Kook, Seunghyi; Kaoud, Tamer S et al. (2015) Arrestin-3-Dependent Activation of c-Jun N-Terminal Kinases (JNKs). Curr Protoc Pharmacol 68:2.12.1-2.12.26
Gurevich, E V; Gurevich, V V (2015) Beyond traditional pharmacology: new tools and approaches. Br J Pharmacol 172:3229-41
Gurevich, Vsevolod V; Gurevich, Eugenia V (2015) Analyzing the roles of multi-functional proteins in cells: The case of arrestins and GRKs. Crit Rev Biochem Mol Biol 50:440-52
Cleghorn, Whitney M; Branch, Kevin M; Kook, Seunghyi et al. (2015) Arrestins regulate cell spreading and motility via focal adhesion dynamics. Mol Biol Cell 26:622-35

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