Determination of the role of DDX3 a factor involved in mammalian RNAi pathway using an shRNA-expression library.

Determination of the role of DDX3 a factor involved in mammalian RNAi pathway using an shRNA-expression library.
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DOI:
10.1371/journal.pone.0059445
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Miyagishi M
Miyagishi M
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Kasim V;Wu S;Taira K;Miyagishi M

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RNA干扰(RNA interference,RNAi)是由双链RNA(double-stranded RNA,dsRNA)诱导的内源性RNA破坏现象。在RNAi中,dsRNA被加工成小干扰RNA(siRNA),其继而触发靶mRNA的切割。在这里,使用短发夹RNA表达文库,我们鉴定了DEAD-box解旋酶3,DDX 3,作为参与RNAi途径的必需因子,并揭示DDX 3与Ago 2共定位,Ago 2是RNAi途径中切割靶mRNA的必需因子。DDX 3显性失活突变体的实验结果进一步证实了该因子影响RNAi活性。总之,DDX 3的功能是确保哺乳动物RNAi途径。总之,我们的研究结果表明,DDX 3是一个新的关键分子,了解哺乳动物RNAi途径的分子机制。
RNA interference (RNAi) is an endogenous RNA-destruction phenomenon induced by certain double-stranded RNAs (dsRNAs). In RNAi, dsRNAs are processed into small interfering RNAs (siRNAs) which in turn trigger the cleavage of the target mRNA. Here, using a short hairpin RNA-expression library, we identified a DEAD-box helicase 3, DDX3, as an essential factor involved in RNAi pathway and revealed that DDX3 is colocalized with Ago2, an essential factor in RNAi pathway that cleaves target mRNA. Results of experiments with a dominant negative mutant of DDX3 further confirmed that this factor affects the RNAi activity. Together, DDX3 functions to assure mammalian RNAi pathway. Together, our results indicate that DDX3 is a new key molecule to understand the molecular mechanism underlying RNAi pathway in mammals.
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