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.

项目摘要/摘要 非细分负链（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，埃博拉病毒）。大多数NNS RNA病毒在细胞质中复制 宿主细胞，而伯恩加病毒1（BODV-1）（一种独特的NNS RNA病毒）在细胞核中复制。 Bodv- 1是动物和人类致命神经系统疾病的病因，尽管在极少数情况下。有趣的是， 在各种基因组中，发现了内源性的伯恩维病毒元素作为古代BONTAVIRASS的化石 包括人类在内的脊椎动物，表明伯恩维病毒与脊椎动物之间存在相互作用 宿主在进化过程中。那就很重要 不仅要了解Bornavires的基本生物学，还要开发针对的治疗靶标 人畜共患潜力。该项目的目的是阐明RNA-的酶促作用 转录中由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蛋白具有酶促活性以进行基因组 转录和复制以及（2）多药P蛋白在维持A L蛋白的转录活性状态。这些假设将通过特定目的测试以阐明 （1）BODV-1 L蛋白在RNA合成和加工中的作用以及（2）P蛋白在形成中 转录活跃的RDRP复合物。在AIM 1中，我们将剖析形成的基础机制 基因组的独特末端和带有BODV-1 L-P复合物的mRNA上的5'-末端盖核结构。 在AIM 2中，我们将解决BODV-1 L-P复合物的3D结构，并研究激活的机理 LNA合成中具有多聚体P蛋白的L蛋白。总的来说，这项研究将推动我们的 了解BODV-1的L蛋白如何进行RNA合成和加工及其共同处理 因子P蛋白。此外，这项研究将揭示RDRP之间的结构相似性和差异 核和细胞质复制NNS RNA病毒的复合物。