Structure and function of Borna disease virus polymerase

博尔纳病病毒聚合酶的结构和功能

• 批准号: 10656952
• 负责人: Tomoaki Ogino
• 金额: $ 24.87万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-17 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656952
• 关键词: Amino Acids; Animals; Baculovirus Expression System; Biochemical; Biochemical Reaction; Bioinformatics; Biological Assay; Biology; Borna Disease; Borna virus; C-terminal; Cell Nucleus; Cells; Complex; Cryoelectron Microscopy; Cytoplasm; Ebola; Elements; Encephalitis; Equus caballus; Evolution; Exhibits; Filovirus; Fossils; Genetic Transcription; Genome; Glycine decarboxylase; Goals; Guanine; Human; In Vitro; Insecta; Interferons; Messenger RNA; Methylation; Methyltransferase; Molecular; Mononegavirales; Mutagenesis; Nuclear; Nuclear Proteins; Paramyxovirus; Play; Pneumovirus; Polymerase; Production; Proteins; RNA Processing; RNA Viruses; RNA biosynthesis; RNA chemical synthesis; RNA-Directed RNA Polymerase; Rabies; Recombinants; Reporting; Rhabdoviridae; Role; Sheep; Squirrel; Structure; System; Testing; Time; Vertebrates; Viral; Viral Genome; Virus; Virus Replication; Zoonoses; cofactor; human pathogen; in vitro Assay; innovation; mRNA capping; nervous system disorder; novel; protein complex; prototype; respiratory; therapeutic target; three dimensional structure; tripolyphosphate; viral RNA

Project Summary/Abstract Non-segmented negative strand (NNS) RNA viruses are highly diversified eukaryotic viruses including significant human pathogens (e.g., rabies, Nipah, Ebola). Most NNS RNA viruses replicate in the cytoplasm of host cells, whereas Borna disease virus 1 (BoDV-1), a unique NNS RNA virus, replicates in the nucleus. BoDV- 1 is a causative agent of fatal neurological diseases in animals and humans, although in rare cases. Interestingly, endogenous bornavirus-like elements were discovered as fossils of ancient bornaviruses in genomes of various vertebrates including humans, indicating that there have been interactions between bornaviruses and vertebrate hosts during evolution. Thus, elucidation of unique strategies of bornaviruses to replicate in host cells is important not only to understand the basic biology of bornaviruses but also to develop therapeutic targets against bornaviruses with zoonotic potential. The goal of this project is to elucidate the enzymatic roles of the RNA- dependent RNA polymerase (RdRp) complex composed of the BoDV-1 L and P proteins in transcription and replication. We hypothesize that (1) the BoDV-1 L protein has enzymatic activities to carry out genome transcription and replication and (2) the multimeric P protein plays structural roles in maintaining a transcriptionally active state of the L protein. These hypotheses will be tested by the specific aims to elucidate the roles of (1) the BoDV-1 L protein in RNA synthesis and processing and (2) the P protein in the formation of a transcriptionally active RdRp complex. In Aim 1, we will dissect the mechanisms underlying the formation of the unique termini of the genome and the 5′-terminal cap core structure on mRNAs with the BoDV-1 L-P complex. In Aim 2, we will solve a 3D structure of the BoDV-1 L-P complex and investigate the mechanism of the activation of the L protein with the multimeric P protein in RNA synthesis. Collectively, this study will advance our understanding of how the L protein of BoDV-1 carries out RNA synthesis and processing together with its co- factor P protein. Furthermore, this study will reveal structural similarities and differences between RdRp complexes of nuclear- and cytoplasmic- replicating NNS RNA viruses.

项目概要/摘要 非分段负链 (NNS) RNA 病毒是高度多样化的真核病毒，包括 重要的人类病原体（例如狂犬病、尼帕病毒、埃博拉病毒）。大多数 NNS RNA 病毒在细胞质中复制 宿主细胞，而博尔纳病病毒 1 (BoDV-1) 是一种独特的 NNS RNA 病毒，在细胞核中复制。博德病毒- 尽管在极少数情况下，1 是动物和人类致命神经系统疾病的病原体。有趣的是， 在各种不同的基因组中发现了内源性博纳病毒样元件作为古代博纳病毒的化石 包括人类在内的脊椎动物，表明博纳病毒和脊椎动物之间存在相互作用 进化过程中的宿主。因此，阐明博纳病毒在宿主细胞中复制的独特策略非常重要 不仅要了解博纳病毒的基本生物学，还要开发针对其的治疗靶点 具有人畜共患潜力的博纳病毒。该项目的目标是阐明 RNA 的酶作用 转录和转录过程中由 BoDV-1 L 和 P 蛋白组成的依赖性 RNA 聚合酶 (RdRp) 复合物 复制。我们假设 (1) BoDV-1 L 蛋白具有进行基因组分析的酶活性 转录和复制以及（2）多聚体 P 蛋白在维持 L蛋白的转录活性状态。这些假设将通过具体目标进行检验以阐明 (1) BoDV-1 L 蛋白在 RNA 合成和加工中的作用以及 (2) P 蛋白在形成 具有转录活性的 RdRp 复合物。在目标 1 中，我们将剖析形成的机制 基因组的独特末端以及具有 BoDV-1 L-P 复合物的 mRNA 上的 5' 末端帽核心结构。 在目标 2 中，我们将解析 BoDV-1 L-P 复合物的 3D 结构并研究激活机制 RNA 合成中 L 蛋白与多聚体 P 蛋白的关系。总的来说，这项研究将推动我们 了解 BoDV-1 的 L 蛋白如何与其共生体一起进行 RNA 合成和加工 P因子蛋白。此外，本研究将揭示 RdRp 之间结构的异同 核和细胞质复制的 NNS RNA 病毒复合体。