Tomari Yukihide, Du Tingting, Haley Benjamin, Schwarz Dianne S, Bennett Ryan, Cook Heather A, Koppetsch Birgit S, Theurkauf William E, Zamore Phillip D
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
Cell. 2004 Mar 19;116(6):831-41. doi: 10.1016/s0092-8674(04)00218-1.
The putative RNA helicase, Armitage (Armi), is required to repress oskar translation in Drosophila oocytes; armi mutant females are sterile and armi mutations disrupt anteroposterior and dorsoventral patterning. Here, we show that armi is required for RNAi. armi mutant male germ cells fail to silence Stellate, a gene regulated endogenously by RNAi, and lysates from armi mutant ovaries are defective for RNAi in vitro. Native gel analysis of protein-siRNA complexes in wild-type and armi mutant ovary lysates suggests that armi mutants support early steps in the RNAi pathway but are defective in the production of active RNA-induced silencing complex (RISC), which mediates target RNA destruction in RNAi. Our results suggest that armi is required for RISC maturation.
假定的RNA解旋酶阿米蒂奇（Armi）是果蝇卵母细胞中抑制osk基因翻译所必需的；armi突变体雌性果蝇不育，且armi突变会破坏前后轴和背腹轴的模式形成。在此，我们表明armi是RNA干扰（RNAi）所必需的。armi突变体雄性生殖细胞无法沉默星状基因（Stellate），该基因是由RNAi内源性调控的基因，并且来自armi突变体卵巢的裂解物在体外RNAi实验中存在缺陷。对野生型和armi突变体卵巢裂解物中蛋白质-小干扰RNA（siRNA）复合物的非变性凝胶分析表明，armi突变体能够支持RNAi途径的早期步骤，但在活性RNA诱导沉默复合体（RISC）的产生方面存在缺陷，RISC在RNAi过程中介导靶RNA的降解。我们的结果表明，RISC成熟需要armi。
