TY - JOUR
T1 - Heterotrimeric Gq proteins act as a switch for GRK5/6 selectivity underlying β-arrestin transducer bias
AU - Kawakami, Kouki
AU - Yanagawa, Masataka
AU - Hiratsuka, Suzune
AU - Yoshida, Misaki
AU - Ono, Yuki
AU - Hiroshima, Michio
AU - Ueda, Masahiro
AU - Aoki, Junken
AU - Sako, Yasushi
AU - Inoue, Asuka
N1 - Funding Information:
We thank Arun K Shukla at Indian Institute of Technology Kanpur for critical discussion and manuscript editing. We also thank Mithu Baidya and Hemlata Agnihotri at Indian Institute of Technology for the Ib30 plasmid; Kayo Sato, Shigeko Nakano and Ayumi Inoue and other members of the laboratory at Tohoku University for their assistance of plasmid preparation and cell-based GPCR assays; Toshifumi Inada and Yoshiro Saito at Tohoku University and Carsten Hoffmann at Friedrich Schiller University Jena for helpful discussion; Hans Bräuner-Osborne at University of Copenhagen for helpful discussion on generation of the GRK-deficient cells. This work was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI grants 17K08264 (A.I.), 21H04791 (A.I.), 21H05113 (A.I.), JPJSBP120213501 (A.I.), JPJSBP120218801 (A.I.) and 20K05760 (Ma.Y.); Grant-in-Aid for JSPS Fellows 19J11256 (K.K.), 20J20669 (Y.O.); Moonshot Research and Development Program JPMJMS2023 (A.I.), PRESTO JPMJPR1331 (A.I.) and JPMJPR20EF (Ma.Y.) from Japan Science and Technology Agency (JST); the PRIME JP19gm5910013 (A.I.), the LEAP JP19gm0010004 (A.I. and J.A.) and the BINDS JP20am0101095 (A.I.) from the Japan Agency for Medical Research and Development (AMED); Daiichi Sankyo Foundation of Life Science (A.I.); The Uehara Memorial Foundation (A.I.).
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - Signaling-biased ligands acting on G-protein-coupled receptors (GPCRs) differentially activate heterotrimeric G proteins and β-arrestins. Although a wealth of structural knowledge about signaling bias at the GPCR level exists (preferential engagement of a specific transducer), little is known about the bias at the transducer level (different functions mediated by a single transducer), partly due to a poor understanding of GPCR kinase (GRK)-mediated GPCR phosphorylation. Here, we reveal a unique role of the Gq heterotrimer as a determinant for GRK-subtype selectivity that regulates subsequent β-arrestin conformation and function. Using the angiotensin II (Ang II) type-1 receptor (AT1R), we show that β-arrestin recruitment depends on both GRK2/3 and GRK5/6 upon binding of Ang II, but solely on GRK5/6 upon binding of the β-arrestin-biased ligand TRV027. With pharmacological inhibition or genetic loss of Gq, GRK-subtype selectivity and β-arrestin functionality by Ang II is shifted to those of TRV027. Single-molecule imaging identifies relocation of AT1R and GRK5, but not GRK2, to an immobile phase under the Gq-inactive, AT1R-stimulated conditions. These findings uncover a previously unappreciated Gq-regulated mechanism that encodes GRK-subtype selectivity and imparts distinct phosphorylation-barcodes directing downstream β-arrestin functions.
AB - Signaling-biased ligands acting on G-protein-coupled receptors (GPCRs) differentially activate heterotrimeric G proteins and β-arrestins. Although a wealth of structural knowledge about signaling bias at the GPCR level exists (preferential engagement of a specific transducer), little is known about the bias at the transducer level (different functions mediated by a single transducer), partly due to a poor understanding of GPCR kinase (GRK)-mediated GPCR phosphorylation. Here, we reveal a unique role of the Gq heterotrimer as a determinant for GRK-subtype selectivity that regulates subsequent β-arrestin conformation and function. Using the angiotensin II (Ang II) type-1 receptor (AT1R), we show that β-arrestin recruitment depends on both GRK2/3 and GRK5/6 upon binding of Ang II, but solely on GRK5/6 upon binding of the β-arrestin-biased ligand TRV027. With pharmacological inhibition or genetic loss of Gq, GRK-subtype selectivity and β-arrestin functionality by Ang II is shifted to those of TRV027. Single-molecule imaging identifies relocation of AT1R and GRK5, but not GRK2, to an immobile phase under the Gq-inactive, AT1R-stimulated conditions. These findings uncover a previously unappreciated Gq-regulated mechanism that encodes GRK-subtype selectivity and imparts distinct phosphorylation-barcodes directing downstream β-arrestin functions.
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U2 - 10.1038/s41467-022-28056-7
DO - 10.1038/s41467-022-28056-7
M3 - Article
C2 - 35078997
AN - SCOPUS:85123463886
SN - 2041-1723
VL - 13
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 487
ER -