EUBACTERIAL ARGONAUTE COMPLEXES BOUND TO GUIDE DNA AND TARGET RNA

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 RNA干扰技术已经彻底改变了我们沉默基因的能力，并作为一种治疗多种疾病的方法有很大的希望。为了最佳地使用这项技术，人们需要破译催化循环中最终导致目标信息降解的各个步骤的分子基础。该项目侧重于降解机制的一个关键组成部分，Argonaute（Ago）蛋白，并解决其在其支架内容纳核酸模板（引导链）的能力，并定义其配对和随后核酸酶介导的信息切割的对齐的关键特征。Ago蛋白由含有PAZ和PIWI的模块组成，作为RNA诱导的沉默复合物（RISC）的关键催化组分，通过介导靶信使RNA（mRNA）的序列特异性切割（包括通过乘客链的初始降解的小干扰RNA（siRNA）的成熟）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期间Ago-核酸复合物的结构，以定义引导链介导的mRNA靶向和降解的潜在机制。

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.