Synergistic and independent actions of multiple terminal nucleotidyl transferases in the 3' tailing of small RNAs in Arabidopsis.
Synergistic and independent actions of multiple terminal nucleotidyl transferases in the 3' tailing of small RNAs in Arabidopsis.
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拟南芥中多个末端核苷酸转移酶在小 RNA 3' 尾部中的协同和独立作用。
DOI：
10.1371/journal.pgen.1005091
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发表时间：
2015-04
期刊：
影响因子：
4.5
通讯作者：
Ren G
中科院分区：
文献类型：
--
作者：
Wang X;Zhang S;Dou Y;Zhang C;Chen X;Yu B;Ren G
植物中的所有类型的小RNA、动物中的piwi相互作用RNA（piRNA）以及果蝇和C.秀丽线虫经历HEN 1介导的3'末端2'-O-甲基化。这种修饰在保护小RNA免于3'尿苷化、修剪和降解中起关键作用。在拟南芥中，HESO 1是尿苷酸化小RNA以触发其降解的主要酶。然而，U-尾仍然存在于空hen 1 heso 1突变体中，表明存在与HESO 1冗余的酶活性。在这里，我们报告，UTP：RNA尿苷酰转移酶（URT 1）是HESO 1的功能paramorphins。URT 1与AGO 1相互作用，当HESO 1不存在时，URT 1在miRNA尿苷酸化中起主要作用。在hen 1 heso 1 urt 1三重突变体中，miRNA的尿苷酸化被全面消除，伴随着3 '至5'修剪的广泛增加。相反，URT 1的破坏似乎不影响异染色质siRNA尿苷酸化。这表明siRNA途径中涉及另外的核苷酸转移酶。对hen 1 heso 1 urt 1三重突变体中的miRNA尾的分析也揭示了先前未知的酶活性的存在，其可以添加非尿苷核苷酸。重要的是，我们发现当HEN 1完全胜任时，HESO 1也可能在miRNA尿苷酸化中与URT 1冗余地起作用。综上所述，我们的数据不仅揭示了HESO 1和URT 1在miRNA的3'尿苷化中的协同作用，而且还揭示了多个末端核苷酸转移酶在小RNA的3'加尾中的独立活性以及尿苷化和修剪之间的拮抗关系。我们的研究结果可能为小RNA 3'端修饰和稳定性控制机制提供进一步的见解。小分子沉默RNA是植物和动物基因表达的关键调节因子。HEN 1介导的3'末端2'-O-甲基化在小RNA稳定性控制中起着至关重要的作用。在缺乏HEN 1的情况下，几种类型的小RNA经常被尿苷化（非模板化尿苷加成）和修剪，这是一种跨物种保守的现象。然而，基本的分子机制几乎不被理解。在这项研究中，我们发现了UTP：RNA尿苷酰转移酶（URT 1），除了在寡聚腺苷酸化mRNA尿苷酸化中发挥作用外，还与HESO 1在miRNA尿苷酸化中发挥协同作用。分析miRNA谱还揭示了在miRNA加尾过程中存在多个末端核苷酸转移酶以及尿苷酸化和修剪之间的拮抗作用。我们相信，这项研究将有助于我们了解各种末端核苷酸转移酶如何识别其底物和协调功能。
All types of small RNAs in plants, piwi-interacting RNAs (piRNAs) in animals and a subset of siRNAs in Drosophila and C. elegans are subject to HEN1 mediated 3’ terminal 2’-O-methylation. This modification plays a pivotal role in protecting small RNAs from 3’ uridylation, trimming and degradation. In Arabidopsis, HESO1 is a major enzyme that uridylates small RNAs to trigger their degradation. However, U-tail is still present in null hen1 heso1 mutants, suggesting the existence of (an) enzymatic activities redundant with HESO1. Here, we report that UTP: RNA uridylyltransferase (URT1) is a functional paralog of HESO1. URT1 interacts with AGO1 and plays a predominant role in miRNA uridylation when HESO1 is absent. Uridylation of miRNA is globally abolished in a hen1 heso1 urt1 triple mutant, accompanied by an extensive increase of 3’-to-5’ trimming. In contrast, disruption of URT1 appears not to affect the heterochromatic siRNA uridylation. This indicates the involvement of additional nucleotidyl transferases in the siRNA pathway. Analysis of miRNA tailings in the hen1 heso1 urt1 triple mutant also reveals the existence of previously unknown enzymatic activities that can add non-uridine nucleotides. Importantly, we show HESO1 may also act redundantly with URT1 in miRNA uridylation when HEN1 is fully competent. Taken together, our data not only reveal a synergistic action of HESO1 and URT1 in the 3’ uridylation of miRNAs, but also independent activities of multiple terminal nucleotidyl transferases in the 3’ tailing of small RNAs and an antagonistic relationship between uridylation and trimming. Our results may provide further insight into the mechanisms of small RNA 3’ end modification and stability control. Small silencing RNAs are key regulators of gene expression in both plants and animals. HEN1-mediated 3’ terminal 2’-O-methylation plays a crucial role in small RNA stability control. In the absence of HEN1, several types of small RNAs become frequently uridylated (non-templated uridine addition) and trimmed, a phenomenon that is conserved across species. However, the underlying molecular mechanism is barely understood. In this study, we have discovered UTP: RNA uridylyltransferase (URT1) that acts synergistically with HESO1 in miRNA uridylation, in addition to its role in oligo-adenylated mRNA uridylation. Analyzing the miRNA profiles also reveals the existence of multiple terminal nucleotidyl transferases in the miRNA tailing process and an antagonistic action between uridylation and trimming. We believe this study will shed light on our understanding of how various terminal nucleotidyl transferases recognize their substrates and function coordinately.
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作者：
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影响因子：
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2012-07-31
影响因子：
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