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.

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。在植物、动物和真菌中，RNase iii相关酶的Dicer家族成员加工双链RNA （dsRNA）以启动小RNA介导的基因沉默机制。为了了解秀丽隐杆线虫Dicer （DCR-1）如何在多种不同的沉默机制中发挥作用，我们使用了基于质谱的蛋白质组学方法来鉴定DCR-1相互作用的蛋白。然后，我们生成并表征了相应基因的缺失等位基因。三种小RNA的产生需要相互作用，包括：(1)小干扰RNA (sirna)，来自外源性dsRNA触发器（exo- sirna）；(2)内源性sirna（内源性sirna）；(3)发育调控microRNAs （miRNAs）。一个相互作用物，保守的rna -磷酸酶同源物PIR-1，是处理假定扩增的DCR-1底物所必需的。内切sirna产生所需的相互作用物包括ERI-1和RRF-3，它们的功能缺失会增强RNAi。我们的研究结果首次揭示了Dicer复杂的生化生态位，并表明dcr -1介导的小rna途径之间存在竞争。