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bioRxiv · 10.1101/251769

A Chemogenetic Platform for Spatio-temporal Control of β-arrestin Translocation and Signaling at G protein-Coupled Receptors

Roth, B. L.·Gotoh, Y.·Giguere, P.·Nichols, D.
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Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=61 SRC=\"FIGDIR/small/251769_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (15K):\norg.highwire.dtl.DTLVardef@1e4bf96org.highwire.dtl.DTLVardef@de4937org.highwire.dtl.DTLVardef@198dd33org.highwire.dtl.DTLVardef@e26f46_HPS_FORMAT_FIGEXP M_FIG C_FIG Although ligand-activated GPCRs induce both G-protein and {beta}-arrestin dependent signaling, gaining precise spatio-temporal control of {beta}-arrestin signaling has proven elusive. Here we describe a platform for specifically activating {beta}-arrestin-dependent signaling in situ. The platform, which we have dubbed \"GA-PAIR\" (GPCR/{beta}-Arrestin -Plant protein and Abscisic acid Induced Recruitment), can be controlled by the inert phytochemical S-(+)-abscisic acid (ABA). ABA induces interaction between ABI1 (ABA Insensitive 1) and PYL1 (Pyrabactin Resistance (PYR) 1-Like), two plant proteins with no mammalian counterparts. We fused ABI to the engineered human muscarinic M3 G protein-coupled receptor (hM3Dq) and PYL1 to {beta}-arrestin2. Addition of ABA induced rapid and nearly complete translocation of the PYL-{beta}-arrestin fusion protein and, importantly, induced both ERK and Akt signaling. Photo-uncaging a new photo-caged ABA analogue allowed us to gain relatively precise spatio-temporal control over {beta}-arrestin translocation. Because GA-PAIR facilitates the exclusive activation of endogenous {beta}-arrestin signaling pathways in the absence of a GPCR ligand or G protein, the GA-PAIR system will facilitate deconvoluting GPCR signaling in situ.

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Roth, B. L., Gotoh, Y., Giguere, P., Nichols, D.. 2018-01-22. A Chemogenetic Platform for Spatio-temporal Control of β-arrestin Translocation and Signaling at G protein-Coupled Receptors. https://doi.org/10.1101/251769

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