| 2005 |
Ric-8B interacts with Gαolf (identified by yeast two-hybrid) and potentiates Gαolf-dependent cAMP accumulation in HEK293 cells, demonstrating it functions as a GEF activator in the olfactory signal transduction pathway. |
Yeast two-hybrid, cAMP accumulation assay in HEK293 cells |
The Journal of neuroscience |
Medium |
15829631
|
| 2006 |
Ric-8B promotes functional heterologous expression of odorant receptors (ORs) on the cell surface and enhances accumulation of Gαolf at the cell periphery, suggesting it enables OR coupling to Gαolf. |
Heterologous expression assay, cell surface receptor expression, fluorescence/immunolocalization |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
16754875
|
| 2008 |
Ric-8B physically interacts with Gγ13 (in addition to Gαolf) and co-localizes with Gαolf, Gβ1, and Gγ13 in cilia of olfactory sensory neurons; Ric-8B interaction with Gαolf is nucleotide-dependent, consistent with a GEF role. |
Co-immunoprecipitation, immunofluorescence localization in olfactory cilia, nucleotide-dependent binding assay |
Molecular and cellular neurosciences |
Medium |
18462949
|
| 2010 |
Ric-8B stabilizes Gαs protein by inhibiting its ubiquitination; Ric-8B knockdown reduces Gαs protein (not mRNA), overexpression increases it, and Ric-8B binding to Gαs is required for this protective effect since splicing variants that cannot bind Gαs fail to inhibit ubiquitination. |
siRNA knockdown, overexpression, ubiquitination assay, Gαs protein/mRNA quantification |
The Journal of biological chemistry |
Medium |
20133939
|
| 2011 |
Purified Ric-8B isoforms (full-length and Δ9) act as GDP release factors/GEFs for Gα subunits: Ric-8BFL stimulates GTPγS binding to Gαs, Gαq, Gα13, and Gαolf; elevates Vmax of Gαs GTP hydrolysis; and the reaction is GTP-dependent, requiring near-Km GTP to release the tight Ric-8B·nucleotide-free Gα intermediate. |
In vitro GEF assay (GTPγS binding), Michaelis-Menten kinetics of GTP hydrolysis, Co-IP with endogenous Gαs in HeLa cells |
The Journal of biological chemistry |
High |
21467038
|
| 2011 |
Ric-8B functions as a molecular chaperone for Gα subunit biosynthesis: Ric-8B knockout ES cells show substantially reduced Gαs protein (without reduced mRNA), indicating Ric-8B is required for maintaining steady-state Gαs abundance. |
Ric-8B knockout ES cells derived from blastocysts, Western blot for Gα protein levels, RT-PCR for mRNA levels |
Science signaling |
High |
22114146
|
| 2013 |
Ric-8A folds nascent Gα subunits during translation; in Ric-8A-depleted rabbit reticulocyte lysate, Gαi, Gαq, and Gα13 (but not Gαs) are improperly folded (trypsin-sensitive). In wheat germ extract (no endogenous Ric-8), Gαq forms aggregates unless Ric-8A is supplemented, whereupon it forms a ~100 kDa Ric-8A:Gαq heterodimer that releases functional Gαq-GTPγS upon GTP addition. This demonstrates the molecular chaperoning function of Ric-8 in folding nascent Gα subunits. |
Cell-free translation (rabbit reticulocyte lysate and wheat germ extract), immunodepletion, limited trypsinolysis protection assay, gel filtration, GTPγS binding |
Proceedings of the National Academy of Sciences of the United States of America |
High |
23431197
|
| 2013 |
In cardiac myocytes, Gαq signaling promotes ubiquitination and degradation of Gαs; co-expression of Ric-8B cancels Gαq-induced Gαs ubiquitination and restores cAMP accumulation. In vitro, Gαq competes with Gαs for binding to Ric-8B, defining a crosstalk mechanism between Gq and Gs pathways mediated by Ric-8B. |
Ubiquitination assay in neonatal rat cardiomyocytes, cAMP accumulation assay, in vitro competitive binding assay |
Genes to cells |
Medium |
24134321
|
| 2017 |
Conditional deletion of Ric-8b specifically in olfactory sensory neurons (OMP-Cre) eliminates Gαolf protein expression in the olfactory epithelium, reduces the mature olfactory sensory neuron layer, increases neuronal cell death, and causes impaired olfactory-guided behavior in mice. |
Conditional knockout mouse (OMP-Cre × Ric-8b floxed), Western blot, immunohistochemistry, behavioral testing |
The Journal of neuroscience |
High |
29118104
|
| 2020 |
Ric-8B hypomorphic mutation in mice reduces mTORC2 activity, as shown by decreased phosphorylation of Akt at Ser473 in mutant embryos and in cultured cells with Ric-8B knockdown, revealing an unexpected role for Ric-8B in mTORC2 regulation. |
Ric-8B hypomorphic mouse model, phospho-Akt (Ser473) western blot, siRNA knockdown in cell lines, RNA-seq |
PLoS genetics |
Medium |
32392211
|
| 2023 |
Cryo-EM structures of Ric-8B in complex with Gαs and Gαolf revealed that Ric-8B accommodates the Gα C-terminal α5 helix in a concave pocket formed by its α-helical repeat elements and distinctly contacts an extended loop unique to Gαs/olf proteins, explaining Ric-8B isoform specificity for Gαs/olf versus Ric-8A specificity for Gαi/o, Gα12/13, and Gαq/11. |
Cryo-EM structure determination, thermal stability assays, cell-based Gαolf folding assay |
Structure |
High |
36931277
|
| 2024 |
Pathogenic GNAO1 encephalopathy mutations in Gαo cause neomorphic gain-of-interaction with Ric-8B (normally only responsible for Gαs/olf), redistributing Ric-8B from cytoplasm to Golgi and imbalancing neuronal G protein signaling networks; the strength of Gαo-Ric-8B interaction correlates with disease severity. |
Co-immunoprecipitation, subcellular localization imaging, GTP binding/hydrolysis assays, functional characterization of >80 Gαo mutants |
The Journal of clinical investigation |
Medium |
38874642
|
| 2024 |
Conditional deletion of Ric-8b in adult mouse cardiac tissue leads to severely reduced contractility, cardiac fibrosis, apoptosis, loss of β-adrenergic L-type calcium channel activation, and downregulation of myosin light chain 2 phosphopeptides. FRET-based assays confirmed selective Ric-8b interaction with Gαs, and conditional Gαs (Gnas) deletion produced an equivalent cardiac phenotype. |
Conditional cardiac knockout mouse, echocardiography, histology, RNA-seq, phosphoproteomics, FRET-based interaction assay, parallel Gnas conditional knockout |
The Journal of biological chemistry |
High |
38879012
|
| 2026 |
RIC8B variant 4 (v4), which lacks the C-terminal cradle loop helix (CLH) domain, suppresses odorant-induced cAMP responses, while variant 1 (full-length) supports OR signaling. AlphaFold3 modeling suggests v4 cannot form hydrogen bonds with Gαs via the CLH domain, potentially producing a dominant-negative misfolded Gαs. |
HEK293T heterologous expression, odorant-induced cAMP assay, AlphaFold3 structural prediction |
Bioscience, biotechnology, and biochemistry |
Low |
41665873
|
| 2011 |
The Ric-8B gene promoter is repressed during osteoblast differentiation by the transcription factor C/EBPβ (LAP* isoform) and by the SWI/SNF chromatin-remodeling complex; C/EBPβ knockdown increases endogenous Ric-8B transcription; nucleosome repositioning accompanies repression. |
Promoter-luciferase assay, siRNA knockdown, ChIP, nucleosome positioning assay in osteoblastic cells |
Molecular and cellular biology |
Medium |
21606199
|
| 2016 |
CREB1 binds proximal CRE sites in the human RIC8B gene promoter (confirmed by ChIP in neuroblastoma cells) and positively regulates RIC8B transcriptional activity, while C/EBPβ binds distal C/EBP sites; luciferase assays identified CRE sites as the dominant elements for basal transcriptional activity. |
Luciferase reporter assay, ChIP, protein-DNA interaction (EMSA-type) analysis |
Journal of cellular biochemistry |
Medium |
26729411
|