36. シロイヌナズナ三量体 G タンパク質α-subunit 結合タンパク質のスクリーニングとその解析

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Heterotrimeric G-protein-mediated signal transduction is a highly conserved signaling mechanism in animal and yeast. In this pathway, extracellular signals are transduced to intracellular effectors through 7-transmembrane receptors (G-protein coupled receptor : GPCR) and heterotrimeric G-protein consisting of three subunits; α, β, and γ. Up to now each subunit has been identified in some species of plants, suggesting that plants have similar signaling mechanisms to those of animal. Because these subunits, however, have not been proved to compose heterotrimer and no GPCR have been identified in plants, plant G-proteins may constitute their own signaling mechanism. AtGPA1,an arabidopsis Gα, has been reported to involved in pleiotropic signaling such as plant hormone functions, cell division cycle, light signaling, pathogen signalsing and so on, but its biochemical function has yet to be clarified. In this study, we performed a yeast two-hybrid screening to identify interacting partners. We used the constitutively active form of AtGPA1 (AtGPA1^<Q222L>; with a Q222L amino acid substitution) in addition to the wild-type form AtGPA1^<WT> as baits. QL21,one of the clones selected by interaction with AtGPA1^<Q222L>, encoded a protein kinase and showed interaction with AtGPA1^<Q222L>, but not with AtGPA1^<WT> in yeast two hybrid system. Their in vitro interaction was confirmed by immunoprecipitation using recombinant AtGPA1 and QL21 expressed in E. coli. QL21 transcript was detected in all the following parts examined; flowers, siliques, upper stems, lower stems, leaves, rosette leaves, and roots. This expression pattern was similar to AtGPA1,suggesting their cooperative biochemical function.
植物化学調節学会の論文
2003-10-10

36. シロイヌナズナ三量体 G タンパク質α-subunit 結合タンパク質のスクリーニングとその解析

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