Bidirectional processing of pri-miRNAs with branched terminal loops by Arabidopsis Dicer-like1.

Bidirectional processing of pri-miRNAs with branched terminal loops by Arabidopsis Dicer-like1.
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DOI:
10.1038/nsmb.2646
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发表时间:
2013-09
影响因子:
16.8
通讯作者:
Zhang X
Zhang X
中科院分区:
生物学1区
文献类型:
--
作者:
Zhu H;Zhou Y;Castillo-González C;Lu A;Ge C;Zhao YT;Duan L;Li Z;Axtell MJ;Wang XJ;Zhang X

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MiRNAs起源于具有特征的茎环结构的初级转录本(pri-miRNAs)。功能上的miRNAs需要对pri-miRNAs进行准确的处理。在这里,我们以pri-miR166家族为范式,报道了pri-miRNA末端环在miRNA生物发生中的决定性作用。我们发现pri-miR166中的多分支末端环在体内显著抑制了miR166的表达。与pri-miRNAs的规范处理不同,末端环分支(TLBed)pri-miRNAs可以被类DICER 1(DCL1)复合体双向处理:从碱基到环和从环到碱基,分别导致miRNAs的生产性和流行性处理。在任何一种情况下,DCL1复合体都在距离参考单链环区16-17个碱基对(BP)的地方典型地切割pri-miRNAs。DCL1还通过其解旋酶结构域向内部环调整加工位点。因此,这些结果为了解具有复杂二级结构的pri-miRNAs的处理机制提供了新的见解。
miRNAs originate from primary transcripts (pri-miRNAs) with characteristic stem-loop structures. Accurate processing of pri-miRNAs is required for functional miRNAs. Here, using pri-miR166 family as a paradigm, we report the decisive role of pri-miRNA terminal loops in miRNA biogenesis. We found that multi-branched terminal loops in pri-miR166s substantially suppressed miR166 expression in vivo. Unlike canonical processing of pri-miRNAs, terminal-loop-branched (TLBed) pri-miRNAs can be processed by Dicer-like1 (DCL1) complexes bi-directionally: from base to loop and from loop to base, resulting in productive and abortive processing of miRNAs, respectively. In either case, DCL1 complexes canonically cut pri-miRNAs at a distance of 16-17 base pairs (bp) from a reference single-stranded loop region. DCL1 also adjusts processing sites toward an internal loop through its helicase domain. Thus, these results provide new insight into the poorly understood processing mechanism of pri-miRNAs with complicated secondary structures.
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