Drosophila melanogaster Dis3 N-terminal domains are required for ribonuclease activities, nuclear localization and exosome interactions.
Drosophila melanogaster Dis3 N-terminal domains are required for ribonuclease activities, nuclear localization and exosome interactions.
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果蝇 Dis3 N 末端结构域是核糖核酸酶活性、核定位和外泌体相互作用所必需的。
DOI：
10.1093/nar/gkq295
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发表时间：
2010
影响因子：
14.9
通讯作者：
Andrulis,ErikD
中科院分区：
文献类型：
--
作者：
Mamolen,Megan;Smith,Alexandra;Andrulis,ErikD
真核细胞使用多种途径来调节RNA的产生、定位和稳定性。这些途径中有几个是由核糖核酸酶控制的。必需的核糖核酸酶Dis 3在不同的RNA代谢途径中起着重要作用。尽管在体外和体内对Dis 3酶的一般特性的了解取得了很大进展，但对Dis 3结构域对其活性、亚细胞定位和蛋白质-蛋白质相互作用的贡献知之甚少。为了解决这些缺口，我们构建了一组果蝇Dis 3（dDis 3）突变体，并评估其体外酶活性及其在S2组织培养细胞中的定位和相互作用。我们表明，dDis 3的N-末端是足够的内切核糖核酸酶活性在体外和适当的N-末端结构域的全长多肽的活性是至关重要的。我们发现，dDis 3的N-末端也有助于其亚细胞分布，是必要的和足够的核心外泌体蛋白的相互作用。最后，dDis 3与dRrp 6和dImportin-α3的相互作用独立于核心相互作用，并且通过两个不同的区域发生。总之，我们的数据表明，dDis 3 N-末端是一个动态的和复杂的枢纽RNA代谢和外泌体相互作用。
Eukaryotic cells use numerous pathways to regulate RNA production, localization and stability. Several of these pathways are controlled by ribonucleases. The essential ribonuclease, Dis3, plays important roles in distinct RNA metabolic pathways. Despite much progress in understanding general characteristics of the Dis3 enzymein vitroandin vivo, much less is known about the contributions of Dis3 domains to its activities, subcellular localization and protein–protein interactions. To address these gaps, we constructed a set ofDrosophila melanogasterDis3 (dDis3) mutants and assessed their enzymatic activityin vitroand their localizations and interactions in S2 tissue culture cells. We show that the dDis3 N-terminus is sufficient for endoribonuclease activityin vitroand that proper N-terminal domain structure is critical for activity of the full-length polypeptide. We find that the dDis3 N-terminus also contributes to its subcellular distribution, and is necessary and sufficient for interactions with core exosome proteins. Finally, dDis3 interaction with dRrp6 and dImportin-α3 is independent of core interactions and occurs though two different regions. Taken together, our data suggest that the dDis3 N-terminus is a dynamic and complex hub for RNA metabolism and exosome interactions.