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来源获得了主要资金， 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在病毒基因表达和复制的晚期阶段至关重要。 Rev与未剪接和单剪接HIV mRNA中的Rev反应元件（RRE）结合，并将核输出受体CRM 1招募到这些转录物上，以促进其转运出核。 除了其在病毒mRNA输出中的功能外，Rev还被认为稳定和调节含RRE的mRNA的翻译，并且可能在将HIV基因组靶向质膜以包封在出芽病毒中方面发挥作用。 Rev/RRE功能所需的Rev相互作用宿主细胞蛋白包括RNA解旋酶DDX 1和DDX 3。我们的目的是通过一系列的pull-down分析，以及随后的多维蛋白质鉴定技术（MudPIT）分析和严格的数据筛选，从HeLa细胞的核和胞浆提取物中鉴定出DDX 1和Rev/RRE结合伴侣。

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.