Viral variants: assessing the impact of natural strain variation on the structure and function of virus replication and transcription factors

病毒变异：评估自然毒株变异对病毒复制和转录因子的结构和功能的影响

• 批准号: 2753035
• 金额: --
• 依托单位: University of Sussex
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2753035
• 关键词: Viral; variants; assessing; impact; natural

The prevalent gamma-herpesvirus Epstein-Barr Virus has two main strains (type1 and type2) that vary only in the sequence of two key transcription factors (TFs) that drive pathogenicity (EBNA2 and EBNA3C). Understanding the functional impact of this variation is hampered by a lack of information on the structure of these TFs. The West lab recently showed how EBNA2 sequence variation impacts interactions with host cell transcription repressors (Ponnusamy et al, 2019).New variation in the essential EBV replication factor, EBNA1, was recently identified. This variation lies both within and outside of the essential DNA-binding domain (DBD). The West lab has solved the high-resolution X-ray structures of variant EBNA1 DBDs (unpublished). Additional variation in the N-terminus of EBNA1 co-segregates with DBD variation, but there is no full length structure of EBNA1 to understand the structural or functional basis of these co-associated changes. This project will: 1. Use protein modelling to gain insights into the impact of type 1 and type 2 variation on the structure and function of EBNA2 and EBNA3C and to test predictions using rational mutagenesis and established biochemical and cell-based functional assays. 2. Use variant EBNA1 DBD X-ray structures and protein modelling to uncover the structural basis for co-associated amino acid variation in the DBD. Dimerisation and DNA-binding assays will be used to test predictions. 3. Use protein modelling to predict the full-length structure of EBNA1 to understand the structural basis of co-associated changes in the EBNA1 N-terminus and DBD. Predictions will be tested in functional assays. Impact: increased understanding of the functional impact of viral variation relevant to many aspects of virology and vaccinology and key information on the structural basis of replication and transcription factor function.

流行的γ-疱疹病毒Epstein-Barr病毒有两种主要毒株（1型和2型），它们仅在驱动致病性的两个关键转录因子（TF）（EBNA 2和EBNA 3C）的序列上有所不同。了解这种变化的功能影响是阻碍了缺乏信息的结构，这些TF。West实验室最近展示了EBNA 2序列变异如何影响与宿主细胞转录抑制因子的相互作用（Ponnusamy et al，2019）。最近发现了EBV复制因子EBNA 1的新变异。这种变异存在于必需DNA结合结构域（DBD）的内部和外部。West实验室已经解决了变体EBNA 1 DBD的高分辨率X射线结构（未发表）。EBNA 1的N-末端的额外变异与DBD变异共分离，但没有EBNA 1的全长结构来理解这些共相关变化的结构或功能基础。该项目将：1。使用蛋白质建模来深入了解1型和2型变异对EBNA 2和EBNA 3C结构和功能的影响，并使用合理的诱变和已建立的生物化学和基于细胞的功能测定来测试预测。2.使用变异EBNA 1 DBD X射线结构和蛋白质建模来揭示DBD中共同相关氨基酸变异的结构基础。二聚化和DNA结合试验将用于检测预测。3.使用蛋白质建模来预测EBNA 1的全长结构，以了解EBNA 1 N-末端和DBD中共同相关变化的结构基础。将在功能测定中检测预测值。影响力：增加了对病毒变异的功能影响的理解，这些变异与病毒学和疫苗学的许多方面以及复制和转录因子功能的结构基础的关键信息有关。