CHARACTERIZATION OF PROTEINS ASSOCIATED WITH A MYO2P CONTAINING RNP

与含有 RNP 的 MYO2P 相关的蛋白质的表征

• 批准号: 7420706
• 负责人: MARK MOOSEKER
• 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/7420706
• 关键词: RNA; proteins; social integration

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. Myo2p, a member of the class V myosins in budding yeast, functions in polarity establishment, in secretory vesicle trafficking in vacuolar inheritance, and in nuclear segregation, golgi, and peroxisome inheritance. Although Myo2p is involved in many processes, only two docking protein complexes are known. Myo2p binds to the vacuolar membrane through the -Vac17p-Vac8p complex, and Kar9p serves as the linker between Myo2p and spindle microtubule to allow nuclear segregation. We have several lines of evidence that show that Myo2p is part of an RNA-protein complex (RNP). The goal of this collaboration is to identify novel proteins in this complex, including linkers of Myo2p to its RNA cargo. We have partially purified the Myo2p RNP complex via differential centrifugation experiments. This complex is present in a high speed pellet fraction (P3). The following experiments show that Myo2p is part of an RNP: 1. On sucrose velocity gradients, Myo2p from P3 fractionation shifts dramatically with the addition of RNase. Myo2p does not bind to actively translating ribosomes, as cycloheximide treatment has no effect on the fractionation pattern. The goal of this collaboration is to identify novel proteins in this complex, including linkers of Myo2p to its RNA cargo.

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。Myo2p是出芽酵母中V类肌球蛋白的一员，在极性建立、液泡遗传中的分泌囊泡运输以及核分离、高尔基体和过氧化物酶体遗传中起作用。虽然Myo2p参与了许多过程，但只有两种对接蛋白复合物是已知的。Myo2p通过-Vac17p-Vac8p复合物结合到液泡膜上，Kar9p作为Myo2p和纺锤体微管之间的连接体，实现核分离。我们有几条证据表明Myo2p是rna -蛋白复合物（RNP）的一部分。此次合作的目标是鉴定该复合体中的新蛋白质，包括Myo2p与其RNA货物的连接物。我们通过差速离心实验部分纯化了Myo2p RNP复合物。这种复合物存在于高速颗粒馏分（P3）中。下面的实验表明Myo2p是RNP的一部分：在蔗糖速度梯度上，随着RNase的加入，P3分馏点的Myo2p发生显著变化。Myo2p不与主动翻译核糖体结合，因为环己亚胺处理对分离模式没有影响。此次合作的目标是鉴定该复合体中的新蛋白质，包括Myo2p与其RNA货物的连接物。