The human DiGeorge syndrome critical region gene 8 and its D-melanogaster homolog are required for miRNA biogenesis
The human DiGeorge syndrome critical region gene 8 and its D-melanogaster homolog are required for miRNA biogenesis
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DOI：
10.1016/j.cub.2004.11.001
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发表时间：
2004-12-14
期刊：
影响因子：
9.2
通讯作者：
Tuschl, T
中科院分区：
文献类型：
--
作者：
Landthaler, M;Yalcin, A;Tuschl, T
MicroRNA（miRNAs）代表在植物和动物中发现的小的非编码RNA家族（最近的综述，参见[1-5]）。miRNA以发育和组织特异性方式表达，并调节部分或完全序列互补的mRNA的翻译效率和稳定性。miRNA以逐步过程从嵌入长RNA聚合酶II初级转录物（pri-miRNA）中的双链RNA前体切除[6-10]。Drosha RNase III催化第一次切除事件，即发夹RNA（pre-miRNA）在细胞核中的释放[11-13]，随后将pre-miRNA输出到细胞质[14-16]，并通过Dicer进一步加工成成熟的miRNA [17-22]。在这里，我们描述了人类DGCR 8，迪乔治综合征的关键区域基因8，和它的果蝇同源。我们提供了生物化学和基于细胞的读数来证明DGCR 8对miRNA初级转录物成熟的要求。果蝇和人DGCR 8的RNAi敲低实验导致pri-miRNA的积累;以及pre-miRNA和成熟miRNA的减少。我们的研究结果表明，DGCR 8和Drosha在人类细胞中相互作用，并存在于功能性pri-miRNA加工复合物中。
MicroRNAs (miRNAs) represent a family of small non-coding RNAs that are found in plants and animals (for recent reviews, see [1-5]). miRNAs are expressed in a developmentally and tissue-specific manner and regulate the translational efficiency and stability of partial or fully sequence-complementary mRNAs. miRNAs are excised in a stepwise process from double-stranded RNA precursors that are embedded in long RNA polymerase II primary transcripts (pri-miRNA) [6-10]. Drosha RNase III catalyzes the first excision event, the release in the nucleus [11-13] of a hairpin RNA (pre-miRNA), which is followed by export of the pre-miRNA to the cytoplasm [14-16] and further processing by Dicer to mature miRNAs [17-22]. Here, we characterize the human DGCR8, the DiGeorge syndrome critical region gene 8, and its Drosophila melanogaster homolog. We provide biochemical and cell-based readouts to demonstrate the requirement of DGCR8 for the maturation of miRNA primary transcripts. RNAi knockdown experiments of fly and human DGCR8 resulted in accumulation of pri-miRNAs; and reduction of pre-miRNAs and mature miRNAs. Our results suggest that DGCR8 and Drosha interact in human cells and reside in a functional pri-miRNA processing complex.