The role of Dicer protein partners in the processing of microRNA precursors.
The role of Dicer protein partners in the processing of microRNA precursors.
复制标题
DOI：
10.1371/journal.pone.0028548
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Krzyzosiak WJ
中科院分区：
文献类型：
--
作者：
Koscianska E;Starega-Roslan J;Krzyzosiak WJ
核糖核酸酶的细胞功能之一是将microRNA前体(前miRNAs)加工成成熟的microRNAs(MiRNAs)。人类脱氧核糖核酸与其蛋白质伙伴AGO2、PACT和TRBP合作执行这一功能。这些辅助蛋白在DICER活性中的确切作用仍然知之甚少。在本研究中，我们使用Northern印迹技术来研究RNAi耗尽AGO2、PACT和TRBP后前miRNA的切割效率和特异性。结果表明，任一DICER蛋白对的抑制不仅对miRNA水平有很大影响，而且对前miRNA水平也有很大影响，对DICER切割的特异性影响很小。对体外产生的DICER切割产物的分析表明，当Pre-miRNA单独用重组DICER处理时，存在切割中间产物。在内源Dester对前miRNA的切割过程中，没有观察到这一中间产物。我们证明了AGO2、PACT和TRBP是DICER在细胞中有效发挥功能所必需的，我们认为这些蛋白的作用之一是确保DICER的两个RNaseIII结构域触发的切割更好地同步。
One of the cellular functions of the ribonuclease Dicer is to process microRNA precursors (pre-miRNAs) into mature microRNAs (miRNAs). Human Dicer performs this function in cooperation with its protein partners, AGO2, PACT and TRBP. The exact role of these accessory proteins in Dicer activity is still poorly understood. In this study, we used the northern blotting technique to investigate pre-miRNA cleavage efficiency and specificity after depletion of AGO2, PACT and TRBP by RNAi. The results showed that the inhibition of either Dicer protein partner substantially affected not only miRNA levels but also pre-miRNA levels, and it had a rather minor effect on the specificity of Dicer cleavage. The analysis of the Dicer cleavage products generated in vitro revealed the presence of a cleavage intermediate when pre-miRNA was processed by recombinant Dicer alone. This intermediate was not observed during pre-miRNA cleavage by endogenous Dicer. We demonstrate that AGO2, PACT and TRBP were required for the efficient functioning of Dicer in cells, and we suggest that one of the roles of these proteins is to assure better synchronization of cleavages triggered by two RNase III domains of Dicer.
登录
查看更多内容
影响因子：
--
作者：
Koscianska E;Starega-Roslan J;Czubala K;Krzyzosiak WJ
通讯作者：
Krzyzosiak WJ
影响因子：
64.8
作者：
Chendrimada, TP;Gregory, RI;Shiekhattar, R
通讯作者：
Shiekhattar, R
影响因子：
11.4
作者：
Lee, Y;Hur, I;Kim, VN
通讯作者：
Kim, VN
影响因子：
4.8
作者：
Kok, Kin Hang;Ng, Ming-Him James;Jin, Dong-Yan
通讯作者：
Jin, Dong-Yan
影响因子：
9.2
作者：
Landthaler, M;Yalcin, A;Tuschl, T
通讯作者：
Tuschl, T