Modulation of gene expression by human cytosolic tRNase Z(L) through 5'-half-tRNA.
Modulation of gene expression by human cytosolic tRNase Z(L) through 5'-half-tRNA.
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DOI：
10.1371/journal.pone.0005908
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发表时间：
2009-06-15
期刊：
影响因子：
3.7
通讯作者：
Nashimoto M
中科院分区：
文献类型：
--
作者：
Elbarbary RA;Takaku H;Uchiumi N;Tamiya H;Abe M;Takahashi M;Nishida H;Nashimoto M
一种长型（tRNase ZL）的tRNA 3 ′加工核糖核酸内切酶（tRNase Z或3 ′ tRNase）可以在模拟tRNA 5 ′-半部分的人工小向导RNA（sgRNA）的指导下在任何所需位点切割任何靶RNA。基于这种酶性质，已经开发了基因沉默技术，其中通过将合成的5 ′-half-tRNA引入细胞中可以下调特定的mRNA水平，该合成的5 ′-half-tRNA被设计成与mRNA的一部分形成前体tRNA样复合物。近年来，有报道称5 ′-half-tRNA片段稳定存在于多种细胞中，但对其生理功能的研究却很少。我们很想知道内源性5 ′-half-tRNA是否在细胞中作为tRNase ZL的sgRNA起作用。在这里，我们表明，人类胞质tRNase ZL通过5 ′-half-tRNA调节基因表达。我们发现与tRNase ZL共免疫沉淀的5 ′-half-tRNAGlu主要存在于细胞质中，在体外起sgRNA的作用，并在人肾293细胞中下调含有其靶序列的荧光素酶mRNA的水平。我们还证明了PPM1F mRNA是由5 ′-half-tRNAGlu引导的tRNase ZL的真正靶点之一。此外，DNA微阵列数据表明，tRNase ZL可能参与p53信号通路和细胞凋亡。
A long form (tRNase ZL) of tRNA 3′ processing endoribonuclease (tRNase Z, or 3′ tRNase) can cleave any target RNA at any desired site under the direction of artificial small guide RNA (sgRNA) that mimics a 5′-half portion of tRNA. Based on this enzymatic property, a gene silencing technology has been developed, in which a specific mRNA level can be downregulated by introducing into cells a synthetic 5′-half-tRNA that is designed to form a pre-tRNA-like complex with a part of the mRNA. Recently 5′-half-tRNA fragments have been reported to exist stably in various types of cells, although little is know about their physiological roles. We were curious to know if endogenous 5′-half-tRNA works as sgRNA for tRNase ZL in the cells. Here we show that human cytosolic tRNase ZL modulates gene expression through 5′-half-tRNA. We found that 5′-half-tRNAGlu, which co-immunoprecipitates with tRNase ZL, exists predominantly in the cytoplasm, functions as sgRNA in vitro, and downregulates the level of a luciferase mRNA containing its target sequence in human kidney 293 cells. We also demonstrated that the PPM1F mRNA is one of the genuine targets of tRNase ZL guided by 5′-half-tRNAGlu. Furthermore, the DNA microarray data suggested that tRNase ZL is likely to be involved in the p53 signaling pathway and apoptosis.
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