Liang Wenxing, Li Changbao, Liu Fang, Jiang Hongling, Li Shuyu, Sun Jiaqiang, Wu Xiaoyan, Li Chuanyou
State Key Laboratory of Plant Genomics, National Centre for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
Cell Res. 2009 Mar;19(3):307-16. doi: 10.1038/cr.2008.317.
Messenger RNA (mRNA) turnover in eukaryotic cells begins with shortening of the poly (A) tail at the 3' end, a process called deadenylation. In yeast, the deadenylation reaction is predominantly mediated by CCR4 and CCR4-associated factor 1 (CAF1), two components of the well-characterised protein complex named CCR4-NOT. We report here that AtCAF1a and AtCAF1b, putative Arabidopsis homologs of the yeast CAF1 gene, partially complement the growth defect of the yeast caf1 mutant in the presence of caffeine or at high temperatures. The expression of AtCAF1a and AtCAF1b is induced by multiple stress-related hormones and stimuli. Both AtCAF1a and AtCAF1b show deadenylation activity in vitro and point mutations in the predicted active sites disrupt this activity. T-DNA insertion mutants disrupting the expression of AtCAF1a and/or AtCAF1b are defective in deadenylation of stress-related mRNAs, indicating that the two AtCAF1 proteins are involved in regulated mRNA deadenylation in vivo. Interestingly, the single and double mutants of AtCAF1a and AtCAF1b show reduced expression of pathogenesis-related (PR) genes PR1 and PR2 and are more susceptible to Pseudomonas syringae pv tomato DC3000 (Pst DC3000) infection, whereas transgenic plants over-expressing AtCAF1a show elevated expression of PR1 and PR2 and increased resistance to the same pathogen. Our results suggest roles of the AtCAF1 proteins in regulated mRNA deadenylation and defence responses to pathogen infections.
真核细胞中的信使核糖核酸(mRNA)周转始于3'端多聚(A)尾的缩短,这一过程称为去腺苷酸化。在酵母中,去腺苷酸化反应主要由CCR4和CCR4相关因子1(CAF1)介导,这是名为CCR4-NOT的特征明确的蛋白质复合物的两个组分。我们在此报告,拟南芥中酵母CAF1基因的假定同源物AtCAF1a和AtCAF1b,在存在咖啡因或高温的情况下部分弥补了酵母caf1突变体的生长缺陷。AtCAF1a和AtCAF1b的表达由多种与应激相关的激素和刺激诱导。AtCAF1a和AtCAF1b在体外均显示出去腺苷酸化活性,并且预测活性位点的点突变会破坏这种活性。破坏AtCAF1a和/或AtCAF1b表达的T-DNA插入突变体在与应激相关的mRNA的去腺苷酸化方面存在缺陷,表明这两种AtCAF1蛋白参与体内mRNA去腺苷酸化的调控。有趣的是,AtCAF1a和AtCAF1b的单突变体和双突变体显示病程相关(PR)基因PR1和PR2的表达降低,并且对丁香假单胞菌番茄致病变种DC3000(Pst DC3000)感染更敏感,而过量表达AtCAF1a的转基因植物显示PR1和PR2的表达升高并且对相同病原体的抗性增加。我们的结果表明AtCAF1蛋白在调控mRNA去腺苷酸化以及对病原体感染的防御反应中的作用。