IDENTIFICATION OF DDX1 AND REV/RRE COFACTORS

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。 人类免疫缺陷病毒（HIV）-1 Rev是病毒基因晚期必需的 表达和复制。 Rev绑定到Rev响应元素（RRE）， 未剪接和单剪接的HIV mRNA，并招募核输出受体CRM1， 这些转录本，以促进其运输出细胞核。 除了其功能外， 在病毒mRNA输出中，Rev也被认为稳定和调节 含有RRE的mRNA，并可能在靶向HIV基因组中发挥作用。 用于包封在出芽病毒中的质膜。 Rev相互作用宿主细胞蛋白 Rev/RRE功能所需的RNA解旋酶包括RNA解旋酶DDX 1和DDX 3。 从HeLa细胞核和胞质提取物中鉴定DDX 1和Rev/RRE结合伴侣 细胞通过一系列的下拉测定，并随后分析多维蛋白 识别技术（MudPIT），然后进行严格的数据过滤。

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. Human immunodeficiency virus (HIV)-1 Rev is essential in the late phase of viral gene expression and replication. Rev binds to the Rev response element (RRE) that occurs in unspliced and singly spliced HIV mRNAs, and recruits the nuclear export receptor CRM1 to these transcripts to facilitate their transport out of the nucleus. In addition to its functions in viral mRNA export, Rev also has been suggested to stabilize and regulate translation of RRE-containing mRNAs, and potentially has a role in targeting the HIV genome to the plasma membrane for encapsulation in the budding virus. Rev-interacting host cell proteins that are required for Rev/RRE functions include the RNA helicases DDX1 and DDX3.Our aim is to identify DDX1 and Rev/RRE binding partners from nuclear and cytosolic extracts of HeLa cells by a series of pull-down assays, and subsequent analysis by multidimensional protein identification technology (MudPIT) followed by stringent data filtering.