IDENTIFICATION OF DDX1 AND REV/RRE COFACTORS

DDX1 和 REV/RRE 辅因子的鉴定

• 批准号: 8171227
• 负责人: Larry R. Gerace
• 金额: $ 0.48万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171227
• 关键词: Binding; Biological Assay; Cell Nucleus; Cell membrane; Cells; Computer Retrieval of Information on Scientific Projects Database; DDX1 gene; Data; Funding; Gene Expression; Genome; Grant; HIV; HIV-1; Hela Cells; Institution; Nuclear; Nuclear Export; Phase; Proteins; RNA Helicase; RNA Splicing; Recruitment Activity; Research; Research Personnel; Resources; Response Elements; Role; Series; Source; Technology; Transcript; Translations; United States National Institutes of Health; Viral; Viral Genes; Virus; mRNA Export; receptor

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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 人类免疫缺陷病毒（HIV）-1 REV在病毒基因的后期至关重要 表达和复制。 Rev与发生在 未填充和单独剪接的艾滋病毒mRNA，并将核出口受体CRM1招募到 这些转录本可促进它们从细胞核中运输。 除了其功能 在病毒mRNA导出中，还建议REV稳定和调节翻译 含有RRE的mRNA，并有可能在靶向HIV基因组到 质膜用于萌芽病毒中的封装。 Rev相互作用宿主细胞蛋白 Rev/RRE功能所需的功能包括RNA解旋酶DDX1和DDX3。 是从HELA的核和胞质提取物中识别DDX1和REV/RRE结合伙伴 细胞通过一系列下拉测定，然后通过多维蛋白进行分析 识别技术（MUDPIT），然后进行严格的数据过滤。