Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs

E.M. Pfeil; J. Brands; N. Merten; T. Vögtle; M. Vescovo; U. Rick; I.-M. Albrecht; N. Heycke; K. Kawakami; Y. Ono; F.M. Ngako Kadji; S. Hiratsuka; J. Aoki; F. Häberlein; M. Matthey; J. Garg; S. Hennen; M.-L. Jobin; K. Seier; D. Calebiro; A. Pfeifer; A. Heinemann; D. Wenzel; G.M. König; B. Nieswandt; B.K. Fleischmann; A. Inoue; K. Simon; E. Kostenis

doi:10.1016/j.molcel.2020.10.027

Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs

E.M. Pfeil, J. Brands, N. Merten, T. Vögtle, M. Vescovo, U. Rick, I.-M. Albrecht, N. Heycke, K. Kawakami, Y. Ono, F.M. Ngako Kadji, S. Hiratsuka, J. Aoki, F. Häberlein, M. Matthey, J. Garg, S. Hennen, M.-L. Jobin, K. Seier, D. CalebiroA. Pfeifer, A. Heinemann, D. Wenzel, G.M. König, B. Nieswandt, B.K. Fleischmann, A. Inoue, K. Simon, E. Kostenis

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Abstract

© 2020 Elsevier Inc.Mechanisms that control mobilization of cytosolic calcium [Ca2+]i are key for regulation of numerous eukaryotic cell functions. One such paradigmatic mechanism involves activation of phospholipase Cβ (PLCβ) enzymes by G protein βγ subunits from activated Gαi-Gβγ heterotrimers. Here, we report identification of a master switch to enable this control for PLCβ enzymes in living cells. We find that the Gαi-Gβγ-PLCβ-Ca2+ signaling module is entirely dependent on the presence of active Gαq. If Gαq is pharmacologically inhibited or genetically ablated, Gβγ can bind to PLCβ but does not elicit Ca2+ signals. Removal of an auto-inhibitory linker that occludes the active site of the enzyme is required and sufficient to empower “stand-alone control” of PLCβ by Gβγ. This dependence of Gi-Gβγ-Ca2+ on Gαq places an entire signaling branch of G-protein-coupled receptors (GPCRs) under hierarchical control of Gq and changes our understanding of how Gi-GPCRs trigger [Ca2+]i via PLCβ enzymes. © 2020 Elsevier Inc.Calcium mobilization by Gi-coupled GPCRs has been perceived as a stand-alone signaling mechanism for almost 30 years. Here, Pfeil et al. report the unexpected discovery that this quintessential biological process does not occur without, and totally depends on, the heterotrimeric G protein subunit Gαq in the living cell context.

Original language	English
Pages (from-to)	940-954.e6
Journal	Molecular cell
Volume	80
Issue number	6
DOIs	https://doi.org/10.1016/j.molcel.2020.10.027
Publication status	Published - 17 Dec 2020
Externally published	Yes

Funding

This work was funded by the Deutsche Forschungsgemeinschaft (DFG) (German Research Foundation) grants 214362475/GRK1873/2 and WE 4461/2-1 (to E.K., D.W., and B.F.), KO 1582/10-1 and KO 1582/10-2 (to E.K.), KO 902/17-1 and KO 902/17-2 (to G.M.K.), the PRIME ( JP19gm5910013 to A.I.) and the LEAP ( JP19gm0010004 to A.I. and J.A.) from the Japan Agency for Medical Research and Development (AMED), JSPS KAKENHI grant ( 17K08264 to A.I.) from Japan Society for the Promotion of Science , and the Sonderforschungsbereich/Transregio 166–Project C1 (to D.C.). D.C. is supported by a Wellcome Trust Senior Research Fellowship.

Funders	Funder number
Japan Agency for Medical Research and Development
Wellcome Trust
Deutsche Forschungsgemeinschaft	WE 4461/2-1, JP19gm0010004, KO 1582/10-1, KO 1582/10-2, 214362475/GRK1873/2, KO 902/17-2, JP19gm5910013, KO 902/17-1
Japan Society for the Promotion of Science	17K08264

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10.1016/j.molcel.2020.10.027

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Pfeil, E. M., Brands, J., Merten, N., Vögtle, T., Vescovo, M., Rick, U., Albrecht, I.-M., Heycke, N., Kawakami, K., Ono, Y., Ngako Kadji, F. M., Hiratsuka, S., Aoki, J., Häberlein, F., Matthey, M., Garg, J., Hennen, S., Jobin, M.-L., Seier, K., ... Kostenis, E. (2020). Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs. Molecular cell, 80(6), 940-954.e6. https://doi.org/10.1016/j.molcel.2020.10.027

@article{22657dad0d3a4ed7ab971d2891bc4d33,

title = "Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs",

abstract = "{\textcopyright} 2020 Elsevier Inc.Mechanisms that control mobilization of cytosolic calcium [Ca2+]i are key for regulation of numerous eukaryotic cell functions. One such paradigmatic mechanism involves activation of phospholipase Cβ (PLCβ) enzymes by G protein βγ subunits from activated Gαi-Gβγ heterotrimers. Here, we report identification of a master switch to enable this control for PLCβ enzymes in living cells. We find that the Gαi-Gβγ-PLCβ-Ca2+ signaling module is entirely dependent on the presence of active Gαq. If Gαq is pharmacologically inhibited or genetically ablated, Gβγ can bind to PLCβ but does not elicit Ca2+ signals. Removal of an auto-inhibitory linker that occludes the active site of the enzyme is required and sufficient to empower “stand-alone control” of PLCβ by Gβγ. This dependence of Gi-Gβγ-Ca2+ on Gαq places an entire signaling branch of G-protein-coupled receptors (GPCRs) under hierarchical control of Gq and changes our understanding of how Gi-GPCRs trigger [Ca2+]i via PLCβ enzymes. {\textcopyright} 2020 Elsevier Inc.Calcium mobilization by Gi-coupled GPCRs has been perceived as a stand-alone signaling mechanism for almost 30 years. Here, Pfeil et al. report the unexpected discovery that this quintessential biological process does not occur without, and totally depends on, the heterotrimeric G protein subunit Gαq in the living cell context.",

author = "E.M. Pfeil and J. Brands and N. Merten and T. V{\"o}gtle and M. Vescovo and U. Rick and I.-M. Albrecht and N. Heycke and K. Kawakami and Y. Ono and {Ngako Kadji}, F.M. and S. Hiratsuka and J. Aoki and F. H{\"a}berlein and M. Matthey and J. Garg and S. Hennen and M.-L. Jobin and K. Seier and D. Calebiro and A. Pfeifer and A. Heinemann and D. Wenzel and G.M. K{\"o}nig and B. Nieswandt and B.K. Fleischmann and A. Inoue and K. Simon and E. Kostenis",

year = "2020",

month = dec,

day = "17",

doi = "10.1016/j.molcel.2020.10.027",

language = "English",

volume = "80",

pages = "940--954.e6",

journal = "Molecular cell",

issn = "1097-2765",

publisher = "Cell Press",

number = "6",

}

Pfeil, EM, Brands, J, Merten, N, Vögtle, T, Vescovo, M, Rick, U, Albrecht, I-M, Heycke, N, Kawakami, K, Ono, Y, Ngako Kadji, FM, Hiratsuka, S, Aoki, J, Häberlein, F, Matthey, M, Garg, J, Hennen, S, Jobin, M-L, Seier, K, Calebiro, D, Pfeifer, A, Heinemann, A, Wenzel, D, König, GM, Nieswandt, B, Fleischmann, BK, Inoue, A, Simon, K & Kostenis, E 2020, 'Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs', Molecular cell, vol. 80, no. 6, pp. 940-954.e6. https://doi.org/10.1016/j.molcel.2020.10.027

TY - JOUR

T1 - Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs

AU - Pfeil, E.M.

AU - Brands, J.

AU - Merten, N.

AU - Vögtle, T.

AU - Vescovo, M.

AU - Rick, U.

AU - Albrecht, I.-M.

AU - Heycke, N.

AU - Kawakami, K.

AU - Ono, Y.

AU - Ngako Kadji, F.M.

AU - Hiratsuka, S.

AU - Aoki, J.

AU - Häberlein, F.

AU - Matthey, M.

AU - Garg, J.

AU - Hennen, S.

AU - Jobin, M.-L.

AU - Seier, K.

AU - Calebiro, D.

AU - Pfeifer, A.

AU - Heinemann, A.

AU - Wenzel, D.

AU - König, G.M.

AU - Nieswandt, B.

AU - Fleischmann, B.K.

AU - Inoue, A.

AU - Simon, K.

AU - Kostenis, E.

PY - 2020/12/17

Y1 - 2020/12/17

N2 - © 2020 Elsevier Inc.Mechanisms that control mobilization of cytosolic calcium [Ca2+]i are key for regulation of numerous eukaryotic cell functions. One such paradigmatic mechanism involves activation of phospholipase Cβ (PLCβ) enzymes by G protein βγ subunits from activated Gαi-Gβγ heterotrimers. Here, we report identification of a master switch to enable this control for PLCβ enzymes in living cells. We find that the Gαi-Gβγ-PLCβ-Ca2+ signaling module is entirely dependent on the presence of active Gαq. If Gαq is pharmacologically inhibited or genetically ablated, Gβγ can bind to PLCβ but does not elicit Ca2+ signals. Removal of an auto-inhibitory linker that occludes the active site of the enzyme is required and sufficient to empower “stand-alone control” of PLCβ by Gβγ. This dependence of Gi-Gβγ-Ca2+ on Gαq places an entire signaling branch of G-protein-coupled receptors (GPCRs) under hierarchical control of Gq and changes our understanding of how Gi-GPCRs trigger [Ca2+]i via PLCβ enzymes. © 2020 Elsevier Inc.Calcium mobilization by Gi-coupled GPCRs has been perceived as a stand-alone signaling mechanism for almost 30 years. Here, Pfeil et al. report the unexpected discovery that this quintessential biological process does not occur without, and totally depends on, the heterotrimeric G protein subunit Gαq in the living cell context.

AB - © 2020 Elsevier Inc.Mechanisms that control mobilization of cytosolic calcium [Ca2+]i are key for regulation of numerous eukaryotic cell functions. One such paradigmatic mechanism involves activation of phospholipase Cβ (PLCβ) enzymes by G protein βγ subunits from activated Gαi-Gβγ heterotrimers. Here, we report identification of a master switch to enable this control for PLCβ enzymes in living cells. We find that the Gαi-Gβγ-PLCβ-Ca2+ signaling module is entirely dependent on the presence of active Gαq. If Gαq is pharmacologically inhibited or genetically ablated, Gβγ can bind to PLCβ but does not elicit Ca2+ signals. Removal of an auto-inhibitory linker that occludes the active site of the enzyme is required and sufficient to empower “stand-alone control” of PLCβ by Gβγ. This dependence of Gi-Gβγ-Ca2+ on Gαq places an entire signaling branch of G-protein-coupled receptors (GPCRs) under hierarchical control of Gq and changes our understanding of how Gi-GPCRs trigger [Ca2+]i via PLCβ enzymes. © 2020 Elsevier Inc.Calcium mobilization by Gi-coupled GPCRs has been perceived as a stand-alone signaling mechanism for almost 30 years. Here, Pfeil et al. report the unexpected discovery that this quintessential biological process does not occur without, and totally depends on, the heterotrimeric G protein subunit Gαq in the living cell context.

U2 - 10.1016/j.molcel.2020.10.027

DO - 10.1016/j.molcel.2020.10.027

M3 - Article

SN - 1097-2765

VL - 80

SP - 940-954.e6

JO - Molecular cell

JF - Molecular cell

IS - 6

ER -

Heterotrimeric G Protein Subunit Gαq Is a Master Switch for Gβγ-Mediated Calcium Mobilization by Gi-Coupled GPCRs

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