Bioscience and Biotechnology Center, Nagoya University, Nagoya 464-8601, Japan.
Plant Cell. 2010 Nov;22(11):3589-602. doi: 10.1105/tpc.110.074542. Epub 2010 Nov 23.
To investigate gibberellin (GA) signaling using the rice (Oryza sativa) GA receptor GIBBERELLIN-INSENSITIVE DWARF1 (GID1) mutant gid1-8, we isolated a suppressor mutant, Suppressor of gid1-1 (Sgd-1). Sgd-1 is an intragenic mutant containing the original gid1-8 mutation (L45F) and an additional amino acid substitution (P99S) in the loop region. GID1(P99S) interacts with the rice DELLA protein SLENDER RICE1 (SLR1), even in the absence of GA. Substitution of the 99th Pro with other amino acids revealed that substitution with Ala (P99A) caused the highest level of GA-independent interaction. Physicochemical analysis using surface plasmon resonance revealed that GID1(P99A) has smaller K(a) (association) and K(d) (dissociation) values for GA(4) than does wild-type GID1. This suggests that the GID1(P99A) lid is at least partially closed, resulting in both GA-independent and GA-hypersensitive interactions with SLR1. One of the three Arabidopsis thaliana GID1s, At GID1b, can also interact with DELLA proteins in the absence of GA, so we investigated whether GA-independent interaction of At GID1b depends on a mechanism similar to that of rice GID1(P99A). Substitution of the loop region or a few amino acids of At GID1b with those of At GID1a diminished its GA-independent interaction with GAI while maintaining the GA-dependent interaction. Soybean (Glycine max) and Brassica napus also have GID1s similar to At GID1b, indicating that these unique GID1s occur in various dicots and may have important functions in these plants.
为了利用水稻(Oryza sativa)赤霉素受体 GIBBERELLIN-INSENSITIVE DWARF1(GID1)突变体 gid1-8 研究赤霉素(GA)信号转导,我们分离到一个抑制突变体 Suppressor of gid1-1(Sgd-1)。Sgd-1 是一个内含子突变体,包含原始 gid1-8 突变(L45F)和环区的另一个氨基酸取代(P99S)。即使没有 GA,GID1(P99S)也与水稻 DELLA 蛋白 SLENDER RICE1(SLR1)相互作用。用其他氨基酸取代第 99 位脯氨酸表明,用丙氨酸(P99A)取代会导致最高水平的非 GA 依赖性相互作用。使用表面等离子体共振的物理化学分析表明,GID1(P99A)对 GA4 的 K(a)(结合)和 K(d)(解离)值均小于野生型 GID1。这表明 GID1(P99A)的盖子至少部分关闭,导致与 SLR1 的非 GA 依赖性和 GA 超敏相互作用。拟南芥 3 个 GID1 中的 1 个 At GID1b 在没有 GA 的情况下也可以与 DELLA 蛋白相互作用,因此我们研究了 At GID1b 的非 GA 依赖性相互作用是否依赖于类似于水稻 GID1(P99A)的机制。At GID1b 的环区或少数氨基酸被 At GID1a 的取代会削弱其与 GAI 的非 GA 依赖性相互作用,同时保持 GA 依赖性相互作用。大豆(Glycine max)和油菜(Brassica napus)也有类似于 At GID1b 的 GID1,这表明这些独特的 GID1 存在于各种双子叶植物中,在这些植物中可能具有重要功能。