NMR studies of viral proteins

结构和生物化学的研究，建议检查蛋白质-蛋白质和蛋白质-脂质相互作用参与病毒出芽和病毒成熟。重点是正链RNA病毒，它们要么引起人类疾病，要么与引起人类疾病的病毒密切相关。对于甲病毒、丙型肝炎病毒、风疹病毒和黄热病病毒（最简单的包膜病毒），将通过NMR和其他方法研究出芽过程和核衣壳稳定性。衣壳稳定和成熟的重要结构特征将通过核磁共振劳斯肉瘤病毒（RSV），一种结构更复杂的逆转录病毒。具体目的是通过确定各种RSV蛋白构建体的三维溶液结构，并表征衣壳（CA）蛋白、核衣壳（NC）蛋白、已提出了衣壳-核衣壳核心颗粒和跨膜E2-糖蛋白，并将通过生化和NMR实验进行测试。进一步的研究，以调查核衣壳的稳定性，黄热病病毒（YFV），丙型肝炎病毒（HCV），风疹病毒（RUBV）使用NMR方法结合诱变结果也提出了。该项目中提出的研究结果，加上该项目中其他地方提出的分子遗传学和晶体学结果，将增强我们对病毒组装的结构和物理基础的理解。

Structural and biochemical studies are proposed to examine protein- protein and protein-lipid interactions involved in viral budding and viral maturation. The focus is on the plus-strand RNA viruses, which either cause human disease or are closely related to viruses which cause human disease. The budding process and nucleocapsid stabilization will be investigated by NMR and other methods for alphavirus, hepatitis C virus, rubella virus, and yellow fever virus, the most simple enveloped viruses. Structural features important for capsid stabilization and maturation will be investigated by NMR for Rous sarcoma virus (RSV), a structurally more complex retrovirus. Specific objectives are to understand the importance of the capsid-nucleocapsid region of the RSV Gag protein by determining 3-dimensional solution structures of various RSV protein constructs, and characterizing interdomain interactions for the C-terminal domain of capsid (CA) protein, the nucleocapsid (NC) protein, and the capsid-nucleocapsid core particles and the transmembrane E2- glycoprotein has been proposed and will be tested by biochemical and NMR experiments. Further studies to investigate nucleocapsid stability are also proposed for yellow fever virus (YFV), hepatitis C virus (HCV), and rubella virus (RUBV) using NMR methods combined with mutagenesis results. Results from the studies proposed in this project, coupled with results from molecular genetics and crystallography proposed elsewhere in the Program Project, will enhance our understanding of the structural and physical basis underlying viral assembly.