Department of Bioscience and Biotechnology, Fukui Prefectural University, 4-1-1 Kenjojima, Matsuoka, Eiheiji-Town, Fukui 910-1195, Japan.
Int J Mol Sci. 2018 Nov 14;19(11):3591. doi: 10.3390/ijms19113591.
Heterotrimeric G proteins are important molecules for regulating plant architecture and transmitting external signals to intracellular target proteins in higher plants and mammals. The rice genome contains one canonical α subunit gene (), four extra-large GTP-binding protein genes (XLGs), one canonical β subunit gene ), and five γ subunit genes (tentatively named , , , , and ). encodes the canonical γ subunit; encodes the plant-specific type of γ subunit with additional amino acid residues at the N-terminus; and the remaining three γ subunit genes encode the atypical γ subunits with cysteine abundance at the C-terminus. We aimed to identify the gene product, Gγ3, in rice tissues using the anti-Gγ3 domain antibody. We also analyzed the truncated protein, Gγ3∆Cys, in the mutant, , using the anti-Gγ3 domain antibody. Based on nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, the immunoprecipitated Gγ3 candidates were confirmed to be Gγ3. Similar to α (Gα) and β subunits (Gβ), Gγ3 was enriched in the plasma membrane fraction, and accumulated in the flower tissues. As mutants show the characteristic phenotype in flowers and consequently in seeds, the tissues that accumulated Gγ3 corresponded to the abnormal tissues observed in mutants.
异三聚体 G 蛋白是调节植物结构和将外部信号传递到高等植物和哺乳动物细胞内靶蛋白的重要分子。水稻基因组包含一个典型的α亚基基因()、四个特大 GTP 结合蛋白基因(XLGs)、一个典型的β亚基基因)和五个γ亚基基因(暂命名为、、、、和)。编码典型的γ亚基;编码植物特异性的γ亚基,在 N 端有额外的氨基酸残基;其余三个γ亚基基因编码 C 端富含半胱氨酸的非典型γ亚基。我们使用抗 Gγ3 结构域抗体,旨在鉴定水稻组织中的基因产物 Gγ3。我们还使用抗 Gγ3 结构域抗体分析了突变体中的截短蛋白 Gγ3∆Cys。基于纳升液相色谱-串联质谱(LC-MS/MS)分析,免疫沉淀的 Gγ3 候选物被确认为 Gγ3。与α(Gα)和β亚基(Gβ)相似,Gγ3 富集在质膜部分,并在花组织中积累。由于突变体在花和种子中表现出特征表型,因此积累 Gγ3 的组织与突变体中观察到的异常组织相对应。