NS3 HELICASE

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新陈代谢需要许多依赖于ATP的分子马达蛋白的作用，这些蛋白被认为运输、重塑和解开RNA序列中的二级结构。这些分子马达中的许多都是死盒或密切相关的蛋白质。许多正链RNA病毒，如丙型肝炎病毒(丙型肝炎病毒)，需要这些蛋白质的活性来进行RNA复制。确定这些蛋白质的机制和它们的特定功能对于我们理解病毒的复制和RNA的代谢是非常重要的。非结构蛋白3(NS3)是一种RNA马达蛋白(或解旋酶)，是丙型肝炎病毒复制所必需的。我们的数据支持这样的假设，即NS3在单体和低聚物种之间平衡存在，并且其RNA解离活性随着齐聚程度的增加而增加。我们的数据表明，NS3解旋酶可以在RNA和DNA上形成细丝。我们希望建立一个合作的努力来可视化这些细丝。随附的手稿描述了我们在DNA上形成细丝的一些证据。然而，支持细丝形成的大部分数据尚未公布。我们希望建立这种合作，以便深入研究NS3的蛋白质-蛋白质和蛋白质-DNA相互作用。我们没有电磁方面的经验，但我们知道有很多例子是用这种方法来表征灯丝的形成。

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 metabolism requires the action of numerous ATP-dependent, molecular motor proteins that are believed to transport, remodel, and unwind secondary structures in RNA sequences. Many of these molecular motors are DEAD-box or closely related proteins. Many positive strand RNA viruses, such as the hepatitis C virus (HCV), require the activity of these proteins for RNA replication. Determination of the mechanism of these proteins and their specific functions is of fundamental importance to our understanding of viral replication as well as RNA metabolism in general. Non-structural protein 3 (NS3) is an RNA motor protein (or helicase), that is necessary for HCV replication. Our data supports the hypothesis that NS3 exists in equilibrium between monomeric and oligomeric species, and that its RNA unwinding activity increases with increasing oligomerization. Our data suggest that NS3 helicase can form filaments on RNA and DNA. We wish to establish a collaborative effort to visualize these filaments. The attached manuscripts describes some of our evidence for filament formation on DNA. However, most of the data supporting filament formation is yet to be published. We wish to establish this collaboration in order to thoroughly investigate protein-protien and protein-DNA interactions of NS3. We have no experience in EM, but are aware of numerous examples in which filament formation has been characterized by this method.