Zhang Xiaolian, Wang Ning, Chen Pei, Gao Mengmeng, Liu Juge, Wang Yufeng, Zhao Tuanjie, Li Yan, Gai Junyi
National Key Laboratory of Crop Genetics and Germplasm Enhancement, National Center for Soybean Improvement, Key Laboratory for Biology and Genetic Improvement of Soybean (General, Ministry of Agriculture), Nanjing Agricultural University, Nanjing, Jiangsu, China.
PLoS One. 2014 Nov 3;9(11):e111120. doi: 10.1371/journal.pone.0111120. eCollection 2014.
Ariadne (ARI) subfamily of RBR (Ring Between Ring fingers) proteins have been found as a group of putative E3 ubiquitin ligases containing RING (Really Interesting New Gene) finger domains in fruitfly, mouse, human and Arabidopsis. Recent studies showed several RING-type E3 ubiquitin ligases play important roles in plant response to abiotic stresses, but the function of ARI in plants is largely unknown. In this study, an ariadne-like E3 ubiquitin ligase gene was isolated from soybean, Glycine max (L.) Merr., and designated as GmARI1. It encodes a predicted protein of 586 amino acids with a RBR supra-domain. Subcellular localization studies using Arabidopsis protoplast cells indicated GmARI protein was located in nucleus. The expression of GmARI1 in soybean roots was induced as early as 2-4 h after simulated stress treatments such as aluminum, which coincided with the fact of aluminum toxicity firstly and mainly acting on plant roots. In vitro ubiquitination assay showed GmARI1 protein has E3 ligase activity. Overexpression of GmARI1 significantly enhanced the aluminum tolerance of transgenic Arabidopsis. These findings suggest that GmARI1 encodes a RBR type E3 ligase, which may play important roles in plant tolerance to aluminum stress.
RBR(Ring Between Ring fingers)蛋白的Ariadne(ARI)亚家族在果蝇、小鼠、人类和拟南芥中被发现是一组含有RING(Really Interesting New Gene)指结构域的推定E3泛素连接酶。最近的研究表明,几种RING型E3泛素连接酶在植物对非生物胁迫的响应中发挥重要作用,但ARI在植物中的功能在很大程度上尚不清楚。在本研究中,从大豆(Glycine max (L.) Merr.)中分离出一个类ariadne E3泛素连接酶基因,并将其命名为GmARI1。它编码一个预测的含有RBR超结构域的586个氨基酸的蛋白质。使用拟南芥原生质体细胞进行的亚细胞定位研究表明,GmARI蛋白位于细胞核中。在模拟胁迫处理(如铝处理)后2-4小时内,大豆根中GmARI1的表达就被诱导,这与铝毒性首先且主要作用于植物根部这一事实相符。体外泛素化试验表明,GmARI1蛋白具有E3连接酶活性。GmARI1的过表达显著增强了转基因拟南芥对铝的耐受性。这些发现表明,GmARI1编码一种RBR型E3连接酶,其可能在植物对铝胁迫的耐受性中发挥重要作用。