Transfer RNA modifications and genes for modifying enzymes in Arabidopsis thaliana.
Transfer RNA modifications and genes for modifying enzymes in Arabidopsis thaliana.
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DOI：
10.1186/1471-2229-10-201
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发表时间：
2010-09-14
影响因子：
5.3
通讯作者：
Zheng B
中科院分区：
文献类型：
--
作者：
Chen P;Jäger G;Zheng B
在生命的所有领域，转移RNA(TRNA)分子都含有修饰过的核苷。TRNA的修饰会影响其编码能力，并影响密码子-反密码子的相互作用。核苷修饰缺陷具有一系列不同的影响，从细菌的毒力降低，人类的神经系统疾病，到植物的基因表达和胁迫反应的变化。本研究的目的是在模式植物拟南芥中寻找与tRNA修饰相关的基因，以了解核苷修饰在植物生长发育中的作用。在这项研究中，我们建立了一种分析模式植物拟南芥和杂交杨树tRNA中修饰核苷的方法。在两个物种的tRNAs中都发现了21个修饰的核苷。为了确定负责植物tRNA修饰的基因，我们对拟南芥基因组进行了候选基因的全局分析。根据酿酒酵母和大肠杆菌同源物的保守结构域，在拟南芥基因组中预测了90多个编码tRNA修饰酶的基因。研究了拟南芥中这些基因的转录积累模式以及这些基因在不同植物物种中的系统发育分布。大多数拟南芥候选基因的转录本在莲座叶和茎尖中含量最高。虽然在拟南芥基因组中发现的大多数tRNA修饰基因家族都存在于其他植物中，但每个家族存在的基因数量存在很大的差异。通过功能缺失突变研究，我们确定了5个tRNA修饰基因(AtTRM10、AtTRM11、AtTRM82、AtKTI12和AtELP1)，它们分别负责4个特定的修饰核苷(m1G、m2G、m7G和ncm5U)(两个基因：AtKTI12和AtELP1分别与ncm5U修饰有关)。AtTRM11突变体表现为早花表型，而AtELP1突变体叶片狭窄，根系生长减慢，角果形状异常，次生枝代有缺陷。利用反向遗传学方法，我们成功地分离和鉴定了拟南芥中的5个tRNA修饰基因。我们的结论是，本研究建立的方法将有助于在多种植物中鉴定tRNA修饰基因。
In all domains of life, transfer RNA (tRNA) molecules contain modified nucleosides. Modifications to tRNAs affect their coding capacity and influence codon-anticodon interactions. Nucleoside modification deficiencies have a diverse range of effects, from decreased virulence in bacteria, neural system disease in human, and gene expression and stress response changes in plants. The purpose of this study was to identify genes involved in tRNA modification in the model plant Arabidopsis thaliana, to understand the function of nucleoside modifications in plant growth and development. In this study, we established a method for analyzing modified nucleosides in tRNAs from the model plant species, Arabidopsis thaliana and hybrid aspen (Populus tremula × tremuloides). 21 modified nucleosides in tRNAs were identified in both species. To identify the genes responsible for the plant tRNA modifications, we performed global analysis of the Arabidopsis genome for candidate genes. Based on the conserved domains of homologs in Sacccharomyces cerevisiae and Escherichia coli, more than 90 genes were predicted to encode tRNA modifying enzymes in the Arabidopsis genome. Transcript accumulation patterns for the genes in Arabidopsis and the phylogenetic distribution of the genes among different plant species were investigated. Transcripts for the majority of the Arabidopsis candidate genes were found to be most abundant in rosette leaves and shoot apices. Whereas most of the tRNA modifying gene families identified in the Arabidopsis genome was found to be present in other plant species, there was a big variation in the number of genes present for each family. Through a loss of function mutagenesis study, we identified five tRNA modification genes (AtTRM10, AtTRM11, AtTRM82, AtKTI12 and AtELP1) responsible for four specific modified nucleosides (m1G, m2G, m7G and ncm5U), respectively (two genes: AtKTI12 and AtELP1 identified for ncm5U modification). The AtTRM11 mutant exhibited an early-flowering phenotype, and the AtELP1 mutant had narrow leaves, reduced root growth, an aberrant silique shape and defects in the generation of secondary shoots. Using a reverse genetics approach, we successfully isolated and identified five tRNA modification genes in Arabidopsis thaliana. We conclude that the method established in this study will facilitate the identification of tRNA modification genes in a wide variety of plant species.
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影响因子：
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