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<p>G protein-coupled receptors (GPCRs) are the largest family of transmembrane receptors in eukaryotes which transduce extracellular stimuli into cells via cytoplasmic trimeric G proteins composed of α, β, and γ subunits. In filamentous fungi, GPCRs are known to be involved in a wide range of physiological functions, including hyphal growth, sexual development, sclerotia formation, and biosynthesis of secondary metabolites, but the interaction specificity between GPCRs and G protein α subunits (Gαs) is largely unknown. In this study, we aimed to comprehensively analyze GPCR-Gα interactions in <i>Aspergillus oryzae</i>. Bimolecular fluorescence complementation (BiFC) assay demonstrated 12 different GPCR-Gα interactions. The negative control experiments ensured that these interactions were true positives. To gain insight into the functional roles of signals generated through GPCR-Gα interactions, we compared the phenotypes of Δ<i>gprM</i>, Δ<i>gprO</i>, and Δ<i>gpaA</i> mutants, since GprM and GprO were shown to interact with GpaA. The results showed that the deletion of <i>gprM</i> and <i>gprO</i> partially reproduced the phenotypes of the Δ<i>gpaA</i> mutant, suggesting that GprM and GprO transmit signals through GpaA. Our results show that the BiFC assay could serve as a tool to predict GPCR-Gα interactions and analyze their functional roles.</p>

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Comprehensive analysis of interactions between G protein-coupled receptors and G protein α subunits in Aspergillus oryzae

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年9月3日

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  • Itsuki Sakamoto,
  • Dong Min Kim,
  • Manabu Arioka

摘要

G protein-coupled receptors (GPCRs) are the largest family of transmembrane receptors in eukaryotes which transduce extracellular stimuli into cells via cytoplasmic trimeric G proteins composed of α, β, and γ subunits. In filamentous fungi, GPCRs are known to be involved in a wide range of physiological functions, including hyphal growth, sexual development, sclerotia formation, and biosynthesis of secondary metabolites, but the interaction specificity between GPCRs and G protein α subunits (Gαs) is largely unknown. In this study, we aimed to comprehensively analyze GPCR-Gα interactions in Aspergillus oryzae. Bimolecular fluorescence complementation (BiFC) assay demonstrated 12 different GPCR-Gα interactions. The negative control experiments ensured that these interactions were true positives. To gain insight into the functional roles of signals generated through GPCR-Gα interactions, we compared the phenotypes of ΔgprM, ΔgprO, and ΔgpaA mutants, since GprM and GprO were shown to interact with GpaA. The results showed that the deletion of gprM and gprO partially reproduced the phenotypes of the ΔgpaA mutant, suggesting that GprM and GprO transmit signals through GpaA. Our results show that the BiFC assay could serve as a tool to predict GPCR-Gα interactions and analyze their functional roles.