NS3 HELICASE

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代谢需要许多ATP依赖性分子马达蛋白的作用，这些蛋白被认为是RNA序列中的转运、重塑和解旋二级结构。 许多这些分子马达是死亡盒或密切相关的蛋白质。 许多正链RNA病毒，如丙型肝炎病毒（HCV），需要这些蛋白质的活性进行RNA复制。 确定这些蛋白质的机制及其特定功能对于我们理解病毒复制以及RNA代谢具有根本重要性。非结构蛋白3（NS 3）是一种RNA马达蛋白（或解旋酶），是HCV复制所必需的。 我们的数据支持的假设，NS 3存在于单体和寡聚物种之间的平衡，其RNA解旋活性随着寡聚化的增加而增加。 我们的数据表明，NS 3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，以可视化这些细丝。 所附手稿描述了我们在DNA上形成细丝的一些证据。 然而，大多数支持细丝形成的数据尚未公布。 我们希望建立这种合作，以彻底研究NS 3的蛋白质-蛋白质和蛋白质-DNA相互作用。我们在EM方面没有经验，但知道许多例子，其中细丝形成的特点是这种方法。

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.