Ago-2-mediated slicer activity is essential for anti-flaviviral efficacy of RNAi.
Ago-2-mediated slicer activity is essential for anti-flaviviral efficacy of RNAi.
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DOI：
10.1371/journal.pone.0027551
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Manjunath N
中科院分区：
文献类型：
--
作者：
Chen S;Chahar HS;Abraham S;Wu H;Pierson TC;Wang XA;Manjunath N
RNA干扰可以由完全互补的siRNA或部分互补的miRNA介导。siRNA广泛用于抑制病毒复制，并且已知RISC中完全互补的siRNA结合的Ago-2降解靶RNA。虽然其他缺乏切割活性的argonaute蛋白也可以结合具有si和miRNA结构的寡核苷酸，但它们是否也有助于抗病毒作用尚不完全清楚。我们测试了si和miRNA结构的寡核苷酸在双荧光素酶测定中的靶抑制以及在表达单个Ago蛋白的ES细胞中对登革热和西尼罗病毒复制的抑制。在荧光素酶测定中，完全互补和部分互补的寡核苷酸都有效地抑制了所有单个Ago表达细胞系中的靶标，尽管完全互补的寡核苷酸在Ago-2+细胞中的功效更高。然而，部分互补的寡核苷酸对任何细胞系中的病毒复制没有影响，而完全互补的siRNA在Ago-2表达中高度有效，但在表达其他Ago蛋白的细胞中无效。无论靶序列是位于病毒的编码区还是3′UTR，都会发生这种情况。我们的结论是，Ago-2切片活性是必不可少的siRNA的抗病毒功效和miRNA介导的翻译抑制/转录不稳定是太弱，以抑制大量表达的黄病毒蛋白。
RNA interference can be mediated by fully complementary siRNA or partially complementary miRNA. siRNAs are widely used to suppress viral replication and the fully complementary siRNA bound Ago-2 in the RISC is known to degrade the target RNA. Although other argonaute proteins lacking slicer activity can also bind oligonucleotides with both si and miRNA structures, whether they can also contribute to antiviral effects is not entirely clear. We tested si and miRNA structured oligos for target repression in dual luciferase assays as well as for inhibition of Dengue and West Nile virus replication in ES cells expressing individual Ago proteins. In luciferase assays, both fully complementary and partially complementary oligos effectively repressed their targets in all individual Ago expressing cell lines, although the efficacy with fully complementary oligos was higher in Ago-2+ cells. However, partially complementary oligos had no effect on virus replication in any cell line, while fully complementary siRNAs were highly effective in Ago-2 expressing, but not in cells expressing other Ago proteins. This occurred irrespective of whether the target sequences were located in the coding region or 3′UTR of the virus. We conclude that Ago-2 slicer activity is essential for anti-viral efficacy of siRNAs and miRNA-mediated translational repression/transcript destabilization is too weak to suppress the abundantly expressed flaviviral proteins.
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影响因子：
13.6
作者：
Kim SS;Garg H;Joshi A;Manjunath N
通讯作者：
Manjunath N
影响因子：
64.5
作者：
Hafner M;Landthaler M;Burger L;Khorshid M;Hausser J;Berninger P;Rothballer A;Ascano M Jr;Jungkamp AC;Munschauer M;Ulrich A;Wardle GS;Dewell S;Zavolan M;Tuschl T
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作者：
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通讯作者：
Manjunath N
DOI：
10.1073/pnas.0703348104
发表时间：
2007-05-22
影响因子：
11.1
作者：
Emara, Mohamed M.;Brinton, Margo A.
通讯作者：
Brinton, Margo A.
影响因子：
4.5
作者：
Rehwinkel, J;Behm-Ansmant, I;Izaurralde, E
通讯作者：
Izaurralde, E