Role of TrpRS in tRNATrp Packaging in Avian Retroviruses

TrpRS 在禽逆转录病毒 tRNATrp 包装中的作用

• 批准号: 7093152
• 负责人: KRISTEN M STEWART MAYNARD
• 金额: $ 4.6万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2007-06-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7093152
• 关键词: Rous sarcoma virus; active sites; aminoacid tRNA ligase; aminoacyl tRNA; cell line; gag protein; gel filtration chromatography; gene mutation; host organism interaction; intermolecular interaction; nucleic acid chemical synthesis; postdoctoral investigator; protein structure function; tryptophan; virus assembly; viruslike particle; western blottings

DESCRIPTION (provided by applicant): Retroviruses utilize cellular tRNA molecules as primers for reverse transcription of viral RNA into double stranded DNA. In avian retroviruses, tRNATrp is used as the primer for reverse transcription and is selectively packaged into virions. Preliminary results indicate that avian tryptophanyl-tRNA synthetase (TrpRS) is selectively packaged into Rous sarcoma virus (RSV) virions. This project will further explore protein-tRNA and protein-protein interactions that aid in the selective packaging of tRNATrp in avian retroviruses; specifically, the interaction between avian Gag and human TrpRS in the presence and absence of tRNATrp. These interactions will be studied using a variety of techniques including affinity pull-down assays, gel filtration, and fluorescence polarization (FP). In addition, the role of TrpRS in selective packaging will be closely examined through the development of mutant forms of tRNATrp and TrpRS, and analysis of their effects on tRNA packaging in vivo. The insights gained from the proposed research may be useful in the design of therapeutic agents that target avian retroviruses.

描述（由申请人提供）：逆转录病毒利用细胞tRNA分子作为引物，将病毒RNA逆转录到双链DNA中。在禽逆转录病毒中，TRNATRP用作逆转录的底漆，并选择性地包装成病毒体。初步结果表明，将鸟类色氨酸-TRNA合成酶（TRPRS）选择性地包装到Rous肉瘤病毒（RSV）病毒中。该项目将进一步探索蛋白质-TRNA和蛋白质 - 蛋白质相互作用，这有助于鸟类逆转录病毒中TRNATRP的选择性包装。具体而言，在存在和不存在trnatRP的情况下，禽堵嘴和人类trpr之间的相互作用。这些相互作用将使用多种技术进行研究，包括亲和力下拉测定，凝胶过滤和荧光极化（FP）。此外，将通过开发TRNATRP和TRPR的突变形式以及对它们对体内tRNA包装的影响的分析，对选择性包装的作用进行仔细研究。从拟议的研究中获得的见解可能对针对禽逆转录病毒的治疗剂的设计有用。