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bioRxiv · 10.1101/2023.01.05.522954

A self-inactivating invertebrate opsin with resistance to retinal depletion optically drives biased signaling toward Gβγ-dependent ion channel modulation

Tsukamoto, H.·Kubo, Y.
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Abstract

Animal opsins, light-sensitive G protein-coupled receptors (GPCRs), have been utilized for optogenetic tools to control G protein-dependent signaling pathways. Upon G protein activation, the Ga and G{beta}{gamma} subunits drive different intracellular signaling pathways, leading to complex cellular responses. For some purposes, Ga-, G{beta}{gamma}-dependent signaling needs to be separately modulated, but these responses are simultaneously evoked due to the 1:1 stoichiometry of Ga and G{beta}{gamma}. Nevertheless, we show temporal activation of G protein using a self-inactivating invertebrate opsin, Platynereis c-opsin1, drives biased signaling for G{beta}{gamma}-dependent GIRK channel activation in a light-dependent manner by utilizing the kinetic difference between G{beta}{gamma}-dependent and Ga-dependent responses. The opsin-induced transient Gi/o activation preferably causes activation of the kinetically-fast G{beta}{gamma}-dependent GIRK channels rather than slower Gi/o-dependent adenylyl cyclase inhibition. Although similar G{beta}{gamma}-biased signaling properties were observed in a selfinactivating vertebrate visual pigment, Platynereis c-opsin1 needs fewer retinal molecules to evoke cellular responses. Furthermore, the G{beta}{gamma}-biased signaling properties of Platynereis c-opsinl are enhanced by genetically fused with RGS8 protein which accelerates G protein inactivation. The self-inactivating invertebrate opsin and its RGS8-fusion protein can function as optical control tools biased for G{beta}{gamma}-dependent ion channel modulation.

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Tsukamoto, H., Kubo, Y.. 2023-01-06. A self-inactivating invertebrate opsin with resistance to retinal depletion optically drives biased signaling toward Gβγ-dependent ion channel modulation. https://doi.org/10.1101/2023.01.05.522954

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