Ric-8A and Ric-8B are positive regulators of heterotrimeric G protein a subunit function. We have recently defined the cellular action of Ric-8 proteins towards Ga subunits. Ric-8 proteins act as molecular chaperones of nucleotide-free Ga conformations during biosynthetic Ga protein folding. In cells in which Ric-8 genes are deleted, cellular abundances of subsets of Ga subunits were reduced by 95%. Ric-8 proteins also exhibit in vitro guanine nucleotide exchange stimulatory activity towards Ga subunits, and we now believe this to be an in vitro phenomenon stemming from the capacity of Ric-8 to induce a partially folded Ga state that has reduced affinity for GDP. We will develop our hypothesis that Ric-8 proteins are Ga subunit molecular chaperones and investigate the mechanisms by which Ric-8 proteins work with other cellular chaperones including the TRiC/CCT (Chaperonin) complex and HSC70/90 systems to fold individual Ga subunits. We have preliminary evidence that there are complex rules and unique chaperone requirements of individual Ga subunits. We will then test a hypothesis that mouse tissue-specific Ric-8A gene deletion can genetically suppress oncogenic G protein or overactive GPCR-driven cancers. Two mouse melanoma models will be used in which GNAQ/11-Q209L (constitutively-active) alleles are the oncogenic driver mutations of metastatic ocular melanoma, and ectopic mGluR5 expression from a melanocyte promoter elicited robust melanoma. We hypothesize that melanocyte mGluR5 inappropriately activates Gq/11 to promote melanoma. Upon proof-of-concept that Ric-8 deletion will suppress the actions of these oncogenes by reducing Gaq/11 cellular abundances, we will perform screens to identify small molecule inhibitors of Ric-8 and G protein interactions. Our goal is to develop Ric-8 inhibitors as a means to indirectly block G protein disease signaling by reducing the abundance of (mutant) Ga subunits folded by Ric-8. Ric-8 inhibitors may prove efficacious against oncogenic G protein diseases and/or as augmentation therapies for existing GPCR therapeutics. We will also conduct a small project to investigate Ric-8A regulation by phosphorylation events and 14-3-3 binding to Ric-8A.

Public Health Relevance

Ric-8 proteins are newly defined regulators of heterotrimeric G protein a subunit biosynthetic production. G proteins transduce drug, hormone, and neurotransmitter influence into physiological cellular actions. The mechanisms by which Ric-8 proteins aid G protein production will be investigated and means of blocking Ric-8 proteins as a way to reduce G protein abundances in pathophysiological states will be explored.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
2R01GM088242-06
Application #
8757091
Study Section
Molecular and Integrative Signal Transduction Study Section (MIST)
Program Officer
Gaillard, Shawn R
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Rochester
Department
Pharmacology
Type
School of Medicine & Dentistry
DUNS #
City
Rochester
State
NY
Country
United States
Zip Code
14627
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Oner, Sukru Sadik; Maher, Ellen M; Gabay, Meital et al. (2013) Regulation of the G-protein regulatory-Gýýi signaling complex by nonreceptor guanine nucleotide exchange factors. J Biol Chem 288:3003-15
Thomas, Celestine J; Briknarová, Klára; Hilmer, Jonathan K et al. (2011) The nucleotide exchange factor Ric-8A is a chaperone for the conformationally dynamic nucleotide-free state of G?i1. PLoS One 6:e23197
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Woodard, Geoffrey E; Huang, Ning-Na; Cho, Hyeseon et al. (2010) Ric-8A and Gi alpha recruit LGN, NuMA, and dynein to the cell cortex to help orient the mitotic spindle. Mol Cell Biol 30:3519-30