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Resources » Paper

Kumar S et al. (2021) G3 (Bethesda) "The neural G protein Galphao tagged with GFP at an internal loop is functional in C. elegans."

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  • Comments on Kumar S et al. (2021) G3 (Bethesda) "The neural G protein Galphao tagged with GFP at an internal loop is functional in C. elegans." (0)

  • Overview

    PMID:
    34003969
    DOI:
    10.1093/g3journal/jkab167
    Status:
    Publication type:
    Journal_article
    WormBase ID:
    WBPaper00061624

    Kumar S, Olson AC, & Koelle MR (2021). The neural G protein Galphao tagged with GFP at an internal loop is functional in C. elegans. G3 (Bethesda). doi:10.1093/g3journal/jkab167

    Galphao is the alpha subunit of the major heterotrimeric G protein in neurons and mediates signaling by every known neurotransmitter, yet the signaling mechanisms activated by Galphao remain to be fully elucidated. Genetic analysis in Caenorhabditis elegans has shown that Galphao signaling inhibits neuronal activity and neurotransmitter release, but studies of the molecular mechanisms underlying these effects have been limited by lack of tools to complement genetic studies with other experimental approaches. Here we demonstrate that inserting the green fluorescent protein (GFP) into an internal loop of the Galphao protein results in a tagged protein that is functional in vivo and that facilitates cell biological and biochemical studies of Galphao. Transgenic expression of Galphao-GFP rescues the defects caused by loss of endogenous Galphao in assays of egg laying and locomotion behaviors. Defects in body morphology caused by loss of Go are also rescued by Galphao-GFP. The Galphao-GFP protein is localized to the plasma membrane of neurons, mimicking localization of endogenous Galphao. Using GFP as an epitope tag, Galphao-GFP can be immunoprecipitated from C. elegans lysates to purify Galphao protein complexes. The Galphao-GFP transgene reported in this study enables studies involving in vivo localization and biochemical purification of Galphao to complement the already well-developed genetic analysis of Galphao signaling.

    We thank Michael Koelle for curating this paper through ACKnowledge (Author Curation to Knowledgebase) 👍

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