Complex G-protein signaling of the adhesion GPCR, ADGRA3.

IF 4 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Sofie M Bagger, Hannes Schihada, Anna L S Walser, Anna Katarzyna Drzazga, Lukas Grätz, Tiago Palmisano, Christina K Kuhn, Maša Mavri, Ann-Sophie Mølleskov-Jensen, Gregory G Tall, Torsten Schöneberg, Signe J Mathiasen, Jonathan A Javitch, Gunnar Schulte, Katja Spiess, Mette M Rosenkilde
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引用次数: 0

Abstract

ADGRA3 (GPR125) is an orphan adhesion G protein-coupled receptor (aGPCR) that plays a role in planar cell polarity (PCP), primarily through recruitment of the signaling components Dishevelled (DVL) during vertebrate gastrulation, and Discs large homolog 1 (Dlg1), which is implicated in cancer. Limited knowledge exists of the signaling capacity in the canonical GPCR pathways of ADGRA3. Here, we employed a series of human cell line-based signaling assays to gain insight into the heterotrimeric G protein-mediated signaling of ADGRA3. We based the design of ADGRA3 constructs on analyses of transcript variants in publicly available human liver and brain RNA-seq datasets. As cleavage in the GPCR autoproteolysis site (GPS) is a hallmark for many aGPCRs, we generated a truncated ADGRA3 (C-terminal fragment, CTF) corresponding to a potential cleavage at the GPS. We found low-level activation of Gi and Gs by ADGRA3 and slightly more by its CTF. As the N terminus of the CTF constitutes a class-defined tethered agonist known as the stachel peptide, we removed the initial three amino acids of the CTF. This resulted in abrogated G protein-mediated signaling, as observed for other aGPCRs. Due to the central role of ADGRA3 in PCP signaling through DVL recruitment, we investigated the G-protein signaling in absence of DVL1-3 and found it sustained. No transcriptional activation was observed downstream of β-catenin in an assay reporting T-cell factor/lymphoid enhancer factor (TCF/LEF)-mediated transcriptional activity. Collectively, this establishes a classical G protein-mediated signaling for ADGRA3 in addition to its association with components of non-canonical Wnt-signaling pathways.

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Journal of Biological Chemistry
Journal of Biological Chemistry Biochemistry, Genetics and Molecular Biology-Biochemistry
自引率
4.20%
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1233
期刊介绍: The Journal of Biological Chemistry welcomes high-quality science that seeks to elucidate the molecular and cellular basis of biological processes. Papers published in JBC can therefore fall under the umbrellas of not only biological chemistry, chemical biology, or biochemistry, but also allied disciplines such as biophysics, systems biology, RNA biology, immunology, microbiology, neurobiology, epigenetics, computational biology, ’omics, and many more. The outcome of our focus on papers that contribute novel and important mechanistic insights, rather than on a particular topic area, is that JBC is truly a melting pot for scientists across disciplines. In addition, JBC welcomes papers that describe methods that will help scientists push their biochemical inquiries forward and resources that will be of use to the research community.
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