Mechanisms of Action of Novel HIV Rev Co-factors

描述(申请人提供)：HIV Rev蛋白是病毒复制所必需的，它的功能是将未剪接的病毒RNA从细胞核输出到细胞质。REV的一个关键细胞辅助因子是CRM1蛋白(也称为XPO1)。对人类核蛋白复合体数据库的挖掘导致了与CRM1相关的两种蛋白质的鉴定--RBM14和PACS1。SiRNA耗竭实验表明，RBM14和PACS1都是REV激活两种不同的REV报告基因所必需的，但不需要依赖于构成运输元件(CTE)或MMTV REV RNA输出途径的报告载体。任何一种蛋白质的缺失都会降低细胞质中未剪接病毒RNA的水平，但会导致细胞核中未剪接病毒RNA的增加。本研究将探讨RBM14和PACS1在REV功能中的作用机制。这项研究将提供对HIV Rev功能的机械性见解，这可能为HIV感染的新治疗策略提供信息。此外，拟议的工作可能有助于深入了解RNA输出的一般机制，这可能与其他人类疾病相关。

DESCRIPTION (provided by applicant): The HIV Rev protein is essential for viral replication and it functions to export unspliced viral RNA from the nucleus to the cytoplasm. A key cellular co-factor for Rev is the CRM1 protein (also named XPO1). The mining of a database of human nuclear protein complexes led to the identification of two proteins that associate with CRM1-- RBM14 and PACS1. SiRNA depletions experiments revealed that both RBM14 and PACS1 are required for Rev activation of two different Rev-reporter plasmids, but not for reporter plasmids dependent upon the Constitutive Transport Element (CTE) or MMTV Rev RNA export pathways. Depletion of either protein reduces the level of unspliced viral RNA in the cytoplasm, but leads to an increase in unspliced viral RNA in the nucleus. The proposed research will investigate the mechanistic roles of RBM14 and PACS1 in Rev function. This research will provide mechanistic insight into HIV Rev function, and this may provide information leading to novel therapeutic strategies for HIV infection. Additionally, the proposed work is likely to shed insight into genera mechanisms of RNA export and this may have relevance to other human diseases.