Delannoy Etienne, Le Ret Monique, Faivre-Nitschke Emmanuelle, Estavillo Gonzalo M, Bergdoll Marc, Taylor Nicolas L, Pogson Barry J, Small Ian, Imbault Patrice, Gualberto José M
Australian Research Council Centre of Excellence in Plant Energy Biology, University of Western Australia, Crawley, 6008 WA, Australia.
Plant Cell. 2009 Jul;21(7):2058-71. doi: 10.1105/tpc.109.066654. Epub 2009 Jul 14.
RNA editing changes the coding/decoding information relayed by transcripts via nucleotide insertion, deletion, or conversion. Editing of tRNA anticodons by deamination of adenine to inosine is used both by eukaryotes and prokaryotes to expand the decoding capacity of individual tRNAs. This limits the number of tRNA species required for codon-anticodon recognition. We have identified the Arabidopsis thaliana gene that codes for tRNA adenosine deaminase arginine (TADA), a chloroplast tRNA editing protein specifically required for deamination of chloroplast (cp)-tRNAArg(ACG) to cp-tRNAArg(ICG). Land plant TADAs have a C-terminal domain similar in sequence and predicted structure to prokaryotic tRNA deaminases and also have very long N-terminal extensions of unknown origin and function. Biochemical and mutant complementation studies showed that the C-terminal domain is sufficient for cognate tRNA deamination both in vitro and in planta. Disruption of TADA has profound effects on chloroplast translation efficiency, leading to reduced yields of chloroplast-encoded proteins and impaired photosynthetic function. By contrast, chloroplast transcripts accumulate to levels significantly above those of wild-type plants. Nevertheless, absence of cp-tRNAArg(ICG) is compatible with plant survival, implying that two out of three CGN codon recognition occurs in chloroplasts, though this mechanism is less efficient than wobble pairing.
RNA编辑通过核苷酸插入、缺失或转换改变转录本传递的编码/解码信息。真核生物和原核生物都利用腺嘌呤脱氨为肌苷对tRNA反密码子进行编辑,以扩大单个tRNA的解码能力。这限制了密码子-反密码子识别所需的tRNA种类数量。我们已经鉴定出拟南芥中编码tRNA腺苷脱氨酶精氨酸(TADA)的基因,它是一种叶绿体tRNA编辑蛋白,专门负责将叶绿体(cp)-tRNAArg(ACG)脱氨为cp-tRNAArg(ICG)。陆地植物的TADA具有一个C末端结构域,其序列和预测结构与原核tRNA脱氨酶相似,并且还具有起源和功能未知的非常长的N末端延伸。生化和突变体互补研究表明,C末端结构域在体外和植物体内对于同源tRNA脱氨都是足够的。TADA的破坏对叶绿体翻译效率有深远影响,导致叶绿体编码蛋白产量降低和光合功能受损。相比之下,叶绿体转录本积累到显著高于野生型植物的水平。然而,缺乏cp-tRNAArg(ICG)与植物存活是相容的,这意味着在叶绿体中三个CGN密码子识别中有两个发生,尽管这种机制不如摆动配对有效。