Format

Send to

Choose Destination
Nat Commun. 2018 Jan 23;9(1):341. doi: 10.1038/s41467-017-02661-3.

Lack of beta-arrestin signaling in the absence of active G proteins.

Author information

1
Molecular, Cellular and Pharmacobiology Section, Institute for Pharmaceutical Biology, University of Bonn, Nussallee 6, 53115, Bonn, Germany.
2
Graduate School of Pharmaceutical Science, Tohoku University, Sendai, 980-8578, Japan.
3
PRESTO, Japan Science and Technology Agency (JST), 4-1-8, Honcho, Kawaguchi, 332-0012, Japan.
4
Institute for Molecular Cell Biology, CMB-Center for Molecular Biomedicine, University Hospital Jena, Hans-Knöll-Strasse2, 07745, Jena, Germany.
5
Bio-Imaging-Center/Rudolf-Virchow-Center, Institute of Pharmacology, University of Wuerzburg, Versbacher Str. 9, 97078, Würzburg, Germany.
6
Institute for Pharmaceutical Biology, University of Bonn, Nussallee 6, 53115, Bonn, Germany.
7
Pharmaceutical Biochemistry and Bioanalytics, Institute of Pharmacy, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
8
AMED-CREST, Japan Agency for Medical Research and Development, 1-7-1 Otemachi, Tokyo, 100-0004, Japan.
9
Molecular Signaling Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), Bethesda, MD, 20892, USA.
10
Molecular, Cellular and Pharmacobiology Section, Institute for Pharmaceutical Biology, University of Bonn, Nussallee 6, 53115, Bonn, Germany. kostenis@uni-bonn.de.

Abstract

G protein-independent, arrestin-dependent signaling is a paradigm that broadens the signaling scope of G protein-coupled receptors (GPCRs) beyond G proteins for numerous biological processes. However, arrestin signaling in the collective absence of functional G proteins has never been demonstrated. Here we achieve a state of "zero functional G" at the cellular level using HEK293 cells depleted by CRISPR/Cas9 technology of the Gs/q/12 families of Gα proteins, along with pertussis toxin-mediated inactivation of Gi/o. Together with HEK293 cells lacking β-arrestins ("zero arrestin"), we systematically dissect G protein- from arrestin-driven signaling outcomes for a broad set of GPCRs. We use biochemical, biophysical, label-free whole-cell biosensing and ERK phosphorylation to identify four salient features for all receptors at "zero functional G": arrestin recruitment and internalization, but-unexpectedly-complete failure to activate ERK and whole-cell responses. These findings change our understanding of how GPCRs function and in particular of how they activate ERK1/2.

PMID:
29362459
PMCID:
PMC5780443
DOI:
10.1038/s41467-017-02661-3
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center