Viral disruption of host transcriptome integrity

病毒破坏宿主转录组完整性

基本信息

批准号：
10666992
负责人：
ANGUS WILSON
金额：
$ 32.88万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2028-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10666992
关键词：
Adenosine Antibodies Antiviral Response Biogenesis Biological Process Catalysis Cell Fractionation Cells Chemicals Chromatin Co-Immunoprecipitations Code Complex Coupled Coupling Cytoplasm DNA Polymerase II Elements Ensure Environment Epigenetic Process Excision Exons Frequencies Gene Expression Genes Genetic Transcription Goals Herpes Simplex Infections Herpesvirus 1 Infection Knowledge Location Mass Spectrum Analysis Measures Mediating Messenger RNA Methylation Methyltransferase Modification Molecular Mutagenesis Nuclear Nuclear RNA Nucleic Acids Pathway interactions Pattern Peptide Signal Sequences Poly A Poly(A) Tail Polyadenylation Polyadenylation Pathway Positioning Attribute Predisposition Productivity Protein Biosynthesis RNA RNA Polymerase II RNA Precursors RNA Processing RNA Splicing RNA-Binding Proteins Recording of previous events Reproduction Resources Role Sedimentation process Signal Transduction Site Small Interfering RNA Structure Testing Transcript Translations Untranslated RNA Untranslated Regions Viral Viral Genes Viral Proteins Viral Regulatory Proteins Virus Virus Diseases Virus Replication Work base inhibitor innovation insight mRNA Stability novel therapeutic intervention premature prevent promoter protein complex recruit transcription termination transcriptome transcriptome sequencing viral RNA

项目摘要

Most viral infections invoke major changes in host gene expression as part of a broader strategy to create an optimal environment for viral replication. By suppressing host protein synthesis, viruses repurpose the biosynthetic resources of the host cell to maximize the accumulation of viral proteins or nucleic acids and thus ultimately boost the yield of new infectious progeny. Herpes simplex virus type 1 (HSV-1) provides a prime example of this important ‘host shutoff’ phenomenon. HSV-1 infection is accompanied by widescale disruption of host mRNA biogenesis through dysregulation of transcription by RNA polymerase II coupled with changes in mRNA stability, splicing, 3’-end formation, and transcription termination. Although multiple studies have implicated the essential viral regulatory protein ICP27 in aspects of shutoff by HSV-1, the exact mechanisms and their relative contributions remain to be elucidated. Recently, we demonstrated that HSV-1 infection induces widespread changes in the subcellular localization of host nuclear factors required for the installation, removal and recognition of internal RNA modifications including methylation at the N6-position of adenosine (m6A). This results in global reductions in the internal base modifications present on both host and viral RNAs. Importantly, we identified the essential viral regulatory protein ICP27 as both necessary and sufficient for this striking effect. Internal RNA modifications such as m6A influence many aspects of mRNA and lncRNA biogenesis including recognition of hardwired splicing and cleavage/polyadenylation signals, and m6A also regulates the export, stability and translation of mature mRNAs. We observed that viral gene expression is sensitive to loss of m6A at the beginning of the infection cycle but is less impacted at later times, coincident with reduced RNA modification frequency. Drawing these observations together we propose that the poorly understood ability of ICP27 to broadly disrupt host gene expression is mediated, at least in part, by its ability to redefine the epigenetic landscape of the host transcriptome. To understand this better, we will more fully characterize the impact of ICP27 expression on the host machinery responsible for RNA chemical modification and for 3’-end processing and transcription termination. Recent studies reveal a mechanistic linkage between the sites of m6A placement and use of nearby polyadenylation and signals and we hypothesize that this is exploited by ICP27 to bring about a widescale disruption of transcription termination (DoTT) that preferentially impacts the host transcriptome in HSV-1 infected cells. To understand why 3’-end formation of HSV-1 mRNAs is largely insensitive to ICP27- mediated DoTT, we will profile m6A installation across the viral transcriptome to determine the impact of m6A on cleavage and polyadenylation site usage within viral transcription units.

大多数病毒感染引起了宿主基因表达的重大变化，这是为病毒复制创造最佳环境的更广泛策略的一部分。通过抑制宿主蛋白的合成，病毒复制宿主细胞的生物合成资源以最大程度地提高病毒蛋白或核酸的积累，从而最终提高了新的感染性后代的产量。单纯疱疹病毒1型（HSV-1）提供了这种重要的“宿主关闭”现象的主要例子。 HSV-1感染是通过通过RNA聚合酶II的转录失调以及mRNA稳定性，剪接，3'-End形成和转录终止的变化来实现宿主mRNA生物发生的。尽管多项研究已经在HSV-1的关闭方面实现了必需的病毒调节蛋白ICP27，但确切的机制及其相对贡献尚待阐明。最近，我们证明了HSV-1感染引起了宿主核因子的亚细胞定位的变化，其安装，去除和识别内部RNA修饰（包括N6位置的N6位置）（M6A）（M6A）所需的内部RNA修饰。这导致宿主和病毒RNA上存在的内部基础修饰的全球减少。重要的是，我们确定必需的病毒调节蛋白ICP27是这种引人注目的效果所必需的和足够的。内部RNA修饰（例如M6A）会影响mRNA和LNCRNA生物发生的许多方面，包括识别硬接线剪接和裂解/聚烯基化信号，M6A还调节了成熟mRNA的出口，稳定性和翻译。我们观察到，病毒基因表达对感染周期开始时M6A的丧失敏感，但在以后的时间影响较小，与RNA修饰频率降低一致。将这些观察结果汇总在一起，我们建议，ICP27广泛破坏宿主基因表达的易于理解的能力至少部分地介导了其重新定义宿主转录组的表观遗传景观的能力。为了更好地理解这一点，我们将更充分地表征ICP27表达对负责RNA化学修饰以及3'-End处理和转录终止的宿主机械的影响。最近的研究揭示了M6A放置位点与近乎聚腺苷酸化和信号的使用之间的机械连接，我们假设ICP27探索了这一点，以实现转录终止的广泛破坏（Dott），优选影响HSV-1感染细胞中的宿主转录组。为了理解为什么HSV-1 mRNA的3'末端形成在很大程度上对ICP27介导的Dott不敏感，我们将在病毒转录组中介绍M6A的安装，以确定M6A对病毒转录单元内裂解和聚丙基化位点使用的影响。