Identification of factors involved in target RNA-directed microRNA degradation.
Identification of factors involved in target RNA-directed microRNA degradation.
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DOI：
10.1093/nar/gkw040
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发表时间：
2016-04-07
影响因子：
14.9
通讯作者：
Pfeffer S
中科院分区：
文献类型：
--
作者：
Haas G;Cetin S;Messmer M;Chane-Woon-Ming B;Terenzi O;Chicher J;Kuhn L;Hammann P;Pfeffer S
微小(Mi)RNA控制其靶基因表达的机制现在已被很好地理解。然而，目前还不太清楚miRNAs本身的水平是如何受到监管的。在特定条件下，丰富的、高度互补的靶RNA可以通过核苷酸加成和核外降解的机制引发miRNA的降解。以前观察到的一种这样的机制是在病毒感染期间自然发生的。到目前为止，这种现象的分子细节尚不清楚。我们在这里报道，互补性的程度和miRNA/靶丰度的比率对于小RNA的有效衰变是至关重要的。使用基于生物素化的反义寡核苷酸的蛋白质组学方法，我们开始确定参与靶向介导的miRNA降解的因素。在回收的蛋白质中，我们鉴定了RNA诱导沉默复合体的成员，但也鉴定了RNA修饰和降解酶。我们进一步验证和表征了其中之一的重要性，即Perlman综合征3‘-5’外切酶Dis3l2。我们证明了该蛋白与ArgAerte2相互作用，并在功能上验证了它在通过人工靶点和在小鼠巨细胞病毒感染的背景下靶向降解miRNA中的作用。
The mechanism by which micro (mi)RNAs control their target gene expression is now well understood. It is however less clear how the level of miRNAs themselves is regulated. Under specific conditions, abundant and highly complementary target RNA can trigger miRNA degradation by a mechanism involving nucleotide addition and exonucleolytic degradation. One such mechanism has been previously observed to occur naturally during viral infection. To date, the molecular details of this phenomenon are not known. We report here that both the degree of complementarity and the ratio of miRNA/target abundance are crucial for the efficient decay of the small RNA. Using a proteomic approach based on the transfection of biotinylated antimiRNA oligonucleotides, we set to identify the factors involved in target-mediated miRNA degradation. Among the retrieved proteins, we identified members of the RNA-induced silencing complex, but also RNA modifying and degradation enzymes. We further validate and characterize the importance of one of these, the Perlman Syndrome 3′-5′ exonuclease DIS3L2. We show that this protein interacts with Argonaute 2 and functionally validate its role in target-directed miRNA degradation both by artificial targets and in the context of mouse cytomegalovirus infection.