The study on molecular functions of the novel RNAi factors DRH-3 and E1 which are required to maintain the chromosome integrity in higher eukaryote

本研究旨在阐明两种RNAi因子DRH-3 （Dicer相关解旋酶-3）和E1（一种具有两个Tudor结构域的新蛋白）的分子功能，我最初发现它们在秀丽隐杆线虫的种系形成中是必不可少的。在本研究中，我生成了10个突变蛋白的表达DNA构建体，并利用大肠杆菌蛋白表达系统成功制备了His和gst标记的DRH-3和E1蛋白的天然版本和一些突变版本，并通过不同标记的天然蛋白通过下拉法展示了DRH-3和E1之间的物理相互作用。此外，对这两个基因的遗传学和细胞学观察表明，DRH-3和E1蛋白在细胞核中作用于相同的RNAi途径。

This study has aimed to clarify the molecular function of two RNAi factors DRH-3 (Dicer Related Helicase-3) and E1 (a novel protein with two Tudor domains) that I originally found to be indispensable for germ-line formation in C. elegans. In this study, I generated the DNA constructs for expression of 10 mutated proteins and successfully prepared His- and GST-tagged-native and some of mutated versions of DRH-3 and E1 proteins using the E. coli protein expression system, and showed the physical interaction between DRH-3 and E1 by pull-down method with differently tagged-native proteins. In addition, genetic and cytological observations on both genes have suggested that DRH-3 and E1 proteins act in the same pathway of RNAi in nucleus.