Asian Natural Environmental Science Center, The University of Tokyo, Nishitokyo, Tokyo 188-0002, Japan.
J Exp Bot. 2013 Dec;64(18):5611-21. doi: 10.1093/jxb/ert327. Epub 2013 Oct 5.
Heterotrimeric G-proteins (G-proteins) have been implicated in ubiquitous signalling mechanisms in eukaryotes. In plants, G-proteins modulate hormonal and stress responses and regulate diverse developmental processes. However, the molecular mechanisms of their functions are largely unknown. A yeast two-hybrid screen was performed to identify interacting partners of the Arabidopsis G-protein β subunit AGB1. One of the identified AGB1-interacting proteins is the Arabidopsis adaptor protein AP-3µ. The interaction between AGB1 and AP-3µ was confirmed by an in vitro pull-down assay and bimolecular fluorescence complementation assay. Two ap-3µ T-DNA insertional mutants were found to be hyposensitive to abscisic acid (ABA) during germination and post-germination growth, whereas agb1 mutants were hypersensitive to ABA. During seed germination, agb1/ap-3µ double mutants were more sensitive to ABA than the wild type but less sensitive than agb1 mutants. However, in post-germination growth, the double mutants were as sensitive to ABA as agb1 mutants. These data suggest that AP-3µ positively regulates the ABA responses independently of AGB1 in seed germination, while AP-3µ does require AGB1 to regulate ABA responses during post-germination growth.
三聚体 G 蛋白(G-proteins)在真核生物中普遍存在的信号转导机制中起作用。在植物中,G 蛋白调节激素和应激反应,并调节多种发育过程。然而,它们功能的分子机制在很大程度上是未知的。进行了酵母双杂交筛选,以鉴定拟南芥 G 蛋白β亚基 AGB1 的相互作用伙伴。鉴定出的 AGB1 相互作用蛋白之一是拟南芥衔接蛋白 AP-3µ。通过体外下拉测定和双分子荧光互补测定证实了 AGB1 和 AP-3µ 之间的相互作用。发现两个 ap-3µ T-DNA 插入突变体在萌发和萌发后生长过程中对脱落酸(ABA)的敏感性降低,而 agb1 突变体对 ABA 敏感。在种子萌发过程中,agb1/ap-3µ 双突变体比野生型对 ABA 更敏感,但比 agb1 突变体对 ABA 更不敏感。然而,在萌发后生长过程中,双突变体对 ABA 的敏感性与 agb1 突变体相同。这些数据表明,AP-3µ 在种子萌发过程中独立于 AGB1 正向调节 ABA 反应,而在萌发后生长过程中,AP-3µ 需要 AGB1 来调节 ABA 反应。