Role of a novel human-virus chimeric protein generated by upstream translation and genetic overprinting

通过上游翻译和基因套印产生的新型人病毒嵌合蛋白的作用

• 批准号: 10514635
• 负责人: Ivan Marazzi
• 金额: $ 21.13万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10514635
• 关键词: Affect; Amino Acids; Animals; Biochemical; Case Study; Cell Nucleus; Cells; Chimeric Proteins; Code; Complement; Complex; Containment; Coupled; Data; Disease; Elements; Ensure; Epidemic; Epithelial Cells; Family; Foundations; Future; Generations; Genes; Genetic; Genetic Transcription; Genome; Genome Mappings; Genomics; Goals; Human; Human Genome; Immunology; Infection; Influenza A virus; Initiator Codon; Knock-in; Life; Life Cycle Stages; Macrophage; Messenger RNA; Methods; Molecular; Monoclonal Antibodies; Mus; Mutation; Nucleotides; Open Reading Frames; Pathogenesis; Phenotype; Physiological; Plant Viruses; Point Mutation; Printing; Process; Proteins; Proteomics; Publications; RNA; RNA Viruses; RNA-Directed RNA Polymerase; Role; Seasons; Sequence Analysis; Terminator Codon; Therapeutic; Tracheal Epithelium; Translating; Translational Regulation; Translations; Untranslated Regions; Viral; Viral Genome; Viral Proteins; Virulence; Virus; Virus Diseases; Virus Replication; base; cell type; design; fitness; genetic approach; genome-wide; immune activation; influenzavirus; mRNA Expression; mutant; novel; pandemic disease; pathogen; pathogenic virus; polypeptide; promoter; prophylactic; protein expression; protein function; respiratory pathogen; ribosome profiling; spatiotemporal; tissue culture; transcriptome; transcriptome sequencing; viral RNA; viral genomics; virology; virus genetics

SUMMARY The capacity of a pathogen to overcome host barriers and establish infection is based on the expression of pathogen-derived proteins. To understand how a pathogen antagonizes the host and establishes infection, we need to have a clear understanding of what proteins a pathogen encodes, how they function, and in what manner they contribute to virulence. The current dogma about many life-threatening pathogens is that they encode just a handful of proteins because of their limited genome. RNA viruses, like Influenza A virus (IAV), are a prime example of this paradigm. Based on this, our understanding of virus life cycles, pathogenesis, and therapeutic or prophylactic methods of disease containment are limited to a small set of known proteins encoded by the viral genome. We hypothesized that, as a result of host-virus genetic interaction, RNA viruses could generate chimeric host-virus genes that are translated into proteins during infection. In fact, IAV, and many other highly pathogenic viruses, use short host RNAs to prime viral transcription to generate viral mRNA. Thus, we further hypothesized that start codons within host primer sequences could drive the expression of chimeric human-viral coding sequences, a process that would depend on the translatability of the viral UTR sequences. Our recent publication indicates the existence of this mechanism, which creates human-virus protein chimeras either as extensions of canonical viral proteins or novel polypeptides by genetic overprinting. This idea is supported by evolutionary analysis and functional data in a few preliminary case studies. The goal of this exploratory R21 application is to characterize in detail the genomic context that allows the generation of viral-human proteins along with characterizing, in a physiological manner, the role of a conserved human-virus protein generated by IAV. A combination of reserve genetic approaches will be used to generate viral mutants and to fully characterize their virulence at the cellular and organismal level. By investigating the role of unknown pathogen- derived proteins, this proposal has the potential to establish their importance, elucidate their role during infection, and provide a proof-of-principle study for future virology- immunology- and genomic studies aimed at defining host-virus proteins in the multiple virus in which they can be generated (3 viral families comprising human, other animal and plant viruses).

概括 病原体克服宿主障碍和建立感染的能力是基于 病原体衍生蛋白的表达。了解病原体如何拮抗宿主 并建立感染，我们需要清楚地了解病原体的蛋白质 编码，它们的功能以及它们有助于毒力的方式。电流 关于许多威胁生命的病原体的教条是它们仅编码少数蛋白质 因为它们的基因组有限。 RNA病毒，例如流感病毒（IAV），是一个很好的例子 这个范式。基于此，我们对病毒生命周期，发病机理和 疾病遏制的治疗或预防性方法仅限于一系列已知的 由病毒基因组编码的蛋白质。 我们假设，由于宿主病毒遗传相互作用，RNA病毒可能会产生 在感染过程中转化为蛋白质的嵌合宿主病毒基因。实际上，iav和 许多其他高度致病的病毒，使用短宿主RNA来素来转录到 产生病毒mRNA。因此，我们进一步假设主机底漆中的启动密码子 序列可以驱动嵌合人类病毒编码序列的表达，这一过程 将取决于病毒UTR序列的可翻译性。我们最近的出版物表明 这种机制的存在，从而形成人类病毒蛋白嵌合体 通过遗传覆盖，规范病毒蛋白或新型多肽的扩展。这个想法是 在一些初步案例研究中得到进化分析和功能数据的支持。这 此探索性R21应用的目标是详细表征 允许在生理学中生成病毒人类蛋白并表征 方式是IAV产生的保守人类病毒蛋白的作用。组合 保留遗传方法将用于产生病毒突变体，并充分表征其 细胞和生物水平的毒力。通过研究未知病原体的作用 该提议有可能确定其重要性，阐明其角色 在感染期间，并为未来的病毒学 - 免疫学和 基因组研究旨在定义多种病毒中的宿主病毒蛋白 产生（3个病毒家族，包括人，其他动物和植物病毒）。