Cellular RNA-binding Zinc Finger Proteins in Viral Infection: Understanding the Rules of Engagement

病毒感染中的细胞 RNA 结合锌指蛋白：了解参与规则

基本信息

批准号：
10230687
负责人：
Jennifer Bohn
金额：
$ 6.6万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10230687
关键词：
2019-nCoV ADP ribosylation Amino Acids Antiviral Agents Antiviral Response Binding Proteins Bioinformatics Biological Biological Process Cell Line Cell physiology Cells Clustered Regularly Interspaced Short Palindromic Repeats Consequentialism Coronavirus Coronavirus Infections Coupled Cytokine Signaling DNA DNA Binding Data Data Set Development Explosion Family Family member Future Gene Expression Regulation Genetic Transcription HIV HIV-1 High-Throughput Nucleotide Sequencing Host Defense Immune Immune response Individual Infection Inflammatory Response Infrastructure Integration Host Factors Investigation Knock-out Knowledge Libraries Messenger RNA Methods Microscopy Molecular Murine leukemia virus Organ Play Process Proteins Proteome Proteomics RNA RNA Binding RNA Decay RNA Processing RNA Splicing RNA Viruses RNA-Binding Proteins Regulation Research Retroviridae Ribonucleases Role Signaling Molecule Small Interfering RNA Species Specificity Specificity Structure Surveys Tissues Up-Regulation Viral Proteins Virus Virus Diseases Virus Replication Work Zinc Fingers attenuation base clinically relevant crosslink cytokine endonuclease falls follow-up global health helicase human coronavirus knock-down macrophage member novel pandemic disease pathogen protein function reconstitution recruit screening therapeutically effective transcriptome transcriptomics ubiquitin-protein ligase viral RNA virus host interaction

项目摘要

Project Summary Viruses such as human immunodeficiency virus-1 (HIV-1) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) are the causative pathogens of on-going pandemics. Understanding host-virus interactions is essential to combatting these viruses. Proteomic approaches have identified host proteins that target viral proteins; however, we lack information regarding host factors that target viral RNA (ZAP) or regulate host RNAs (TTP, MCPIP1, and Roquin-1) during infection. Interestingly, the short list of known RNA-binding effectors shares a common CCCH-type zinc finger (ZnF) motif. Unlike other cellular ZnF proteins, most CCCH-type ZnF proteins bind RNA rather than DNA and account for nearly 40% of all RNA-binding ZnF proteins. Furthermore, many of these proteins have poorly annotated functions. I propose that unidentified CCCH ZnF proteins regulate host and viral RNAs during infection. Through systematic siRNA screening, I will identify CCCH-ZnF proteins that modulate the replication of two clinically relevant viruses: human immunodeficiency virus type 1 (HIV-1) and human coronavirus OC43 (HCoV-OC43). I have already identified exciting CCCH-ZnF proteins that have significant beneficial or deleterious effects on HIV-1 and HCoV-OC43 replication. While unique hits were identified for each virus, seven CCCH-ZnF proteins are shared hits in both HIV-1 and HCoV-OC43 screens, suggesting that some of these factors may be broadly antiviral. The proposed research seeks to understand the requirement for RNA-binding by ZnF motif(s) in these host-virus interactions and will use transcriptomics, proteomics, and microscopy approaches to elucidate detailed mechanisms of action. Preliminary results justify this rapid and robust screening method for identification of CCCH-ZnF (and other ZnF-type) factors involved in replication of diverse virus families. Furthermore, I will establish an efficient, streamlined workflow to determine the most promising hits for mechanistic exploration. My work will uncover novel host factors networks that are involved in virus replication and assign molecular functions to cellular CCCH ZnF proteins.

项目概要人类免疫缺陷病毒-1 (HIV-1) 和严重急性呼吸道综合症等病毒冠状病毒-2 (SARS-CoV2) 是当前流行病的致病病原体。了解宿主病毒相互作用对于对抗这些病毒至关重要。蛋白质组学方法已经鉴定出宿主蛋白靶向病毒蛋白；然而，我们缺乏关于靶向病毒 RNA (ZAP) 或调节的宿主因子的信息。感染期间的宿主 RNA（TTP、MCPIP1 和 Roquin-1）。有趣的是，已知 RNA 结合的简短列表效应器具有共同的 CCCH 型锌指 (ZnF) 基序。与其他细胞 ZnF 蛋白不同，大多数 CCCH 型 ZnF 蛋白结合 RNA 而不是 DNA，因此近 40% 的 RNA 结合 ZnF 蛋白。此外，许多这些蛋白质的注释很差功能。我认为未鉴定的 CCCH ZnF 蛋白在感染过程中调节宿主和病毒 RNA。通过系统的 siRNA 筛选，我将鉴定出调节复制的 CCCH-ZnF 蛋白两种临床相关病毒：人类免疫缺陷病毒 1 型 (HIV-1) 和人类冠状病毒 OC43 （HCoV-OC43）。我已经发现了令人兴奋的 CCCH-ZnF 蛋白，它们具有显着的有益或对 HIV-1 和 HCoV-OC43 复制的有害影响。虽然每种病毒都有独特的命中，但有七种病毒 CCCH-ZnF 蛋白在 HIV-1 和 HCoV-OC43 筛查中都是共同命中的，这表明其中一些因素可能具有广泛的抗病毒作用。拟议的研究旨在了解 RNA 结合的要求通过 ZnF 基序在这些宿主-病毒相互作用中进行研究，并将使用转录组学、蛋白质组学和显微镜检查阐明详细作用机制的方法。初步结果证明了这种快速、稳健的 CCCH-ZnF 筛选方法（和其他 ZnF 型）因子参与不同病毒家族的复制。此外，我将建立一个高效、简化工作流程，以确定最有希望进行机械探索的命中。我的工作将揭开参与病毒复制并将分子功能分配给细胞 CCCH 的新型宿主因子网络 ZnF 蛋白质。