EUBACTERIAL ARGONAUTE COMPLEXES BOUND TO GUIDE DNA AND TARGET RNA

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 RNA干扰技术彻底改变了我们沉默基因的能力，并有望成为一种治疗多种疾病的方法。为了以最佳方式使用这项技术，人们需要破译催化循环中最终导致靶向信息降解的各个步骤所基于的分子基础。该项目侧重于降解机制的一个关键组成部分--ArgAerte(AGO)蛋白质，并解决其在其支架内容纳核酸模板(引导链)的能力，并确定其配对和随后的核酸酶介导的消息切割的关键特征。AGO蛋白作为RNA诱导沉默复合体(RISC)的关键催化成分，由含有PAZ和PIWI的模块组成，通过介导靶信使RNA(MRNA)的序列特异性切割，包括通过客体链的初始降解使小干扰RNA(SiRNA)成熟，在RNA干扰途径中发挥核心作用。 我们的目标是确定在RNA干扰催化循环的不同组装和功能步骤中的核酸复合体的结构，以确定引导链介导的靶向和降解mRNA的机制。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. RNA interference techniques have revolutionized our ability to silence genes and hold great promise as a therapeutic approach against a diverse set of diseases. To use this technology optimally, one needs to decipher the molecular basis underlying individual steps in the catalytic cycle that eventually lead to degradation of the targeted message. This project focuses on one critical component of the degradation machinery, the Argonaute (Ago) protein, and addresses its ability to accommodate a nucleic acid template (guide strand) within its scaffold, and defines key features of its alignment for pairing and subsequent nuclease-mediated cleavage of the message. The Ago protein, composed of PAZ- and PIWI-containing modules, as a key catalytic component of the RNA-induced silencing complex (RISC), plays a central role in the RNA interference pathway by mediating sequence-specific cleavage of target messenger RNA (mRNA), including the maturation of small interfering RNA (siRNA) through initial degradation of the passenger strand. Our goal is to determine structures of Ago-nucleic acid complexes during distinct assembly and functional steps of the RNA interference catalytic cycle in order to define the mechanisms underlying guide strand mediated targeting and degradation of mRNA.