FUNCTIONAL PROTEOMICS REVEALS THE BIOCHEMICAL NICHE OF C ELEGANS DCR-1 IN MULT

功能蛋白质组学揭示了秀丽隐杆线虫 DCR-1 在多种动物中的生化利基

• 批准号: 7420805
• 负责人: CRAIG C MELLO
• 金额: $ 0.29万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7420805
• 关键词: RNA

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. In plants, animals, and fungi, members of the Dicer family of RNase III-related enzymes process double-stranded RNA (dsRNA) to initiate small-RNA-mediated gene-silencing mechanisms. To learn how C. elegans Dicer, DCR-1, functions in multiple distinct silencing mechanisms, we used a mass-spectrometry-based proteomics approach to identify DCR-1-interacting proteins. We then generated and characterized deletion alleles for the corresponding genes. The interactors are required for production of three species of small RNA, including (1) small interfering RNAs (siRNAs), derived from exogenous dsRNA triggers (exo-siRNAs); (2) siRNAs derived from endogenous triggers (endo-siRNAs); and (3) developmental regulatory microRNAs (miRNAs). One interactor, the conserved RNA-phosphatase homolog PIR-1, is required for the processing of a putative amplified DCR-1 substrate. Interactors required for endo-siRNA production include ERI-1 and RRF-3, whose loss of function enhances RNAi. Our findings provide a first glimpse at the complex biochemical niche of Dicer and suggest that competition exists between DCR-1-mediated small-RNA pathways.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中出现。列出的机构是中心的机构，不一定是研究者的机构。在植物、动物和真菌中，RNase III 相关酶 Dicer 家族的成员处理双链 RNA (dsRNA) 以启动小 RNA 介导的基因沉默机制。为了了解线虫 Dicer (DCR-1) 如何在多种不同的沉默机制中发挥作用，我们使用基于质谱的蛋白质组学方法来识别 DCR-1 相互作用蛋白。然后我们生成并表征了相应基因的缺失等位基因。相互作用子是产生三种小RNA所必需的，包括（1）源自外源dsRNA触发物（exo-siRNA）的小干扰RNA（siRNA）； (2)源自内源性触发物的siRNA(endo-siRNA)； (3) 发育调节 microRNA (miRNA)。处理假定的扩增 DCR-1 底物需要一种相互作用子，即保守的 RNA 磷酸酶同系物 PIR-1。内切 siRNA 生产所需的相互作用物包括 ERI-1 和 RRF-3，其功能丧失会增强 RNAi。我们的研究结果首次展示了 Dicer 复杂的生化生态位，并表明 DCR-1 介导的小 RNA 通路之间存在竞争。