Roles of Experimentally Evolved Vaccinia Mutations in Antagonizing Cell Defenses

实验进化的痘苗突变在拮抗细胞防御中的作用

基本信息

批准号：
8769973
负责人：
ADAM P. GEBALLE
金额：
$ 22万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-15 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8769973
关键词：
Affect Antiviral Agents Back Biological Assay Biological Models Cells Cercopithecus pygerythrus Conflict (Psychology)Coupled Cytomegalovirus DNA Viruses DNA-Directed RNA Polymerase Defect Detection Double-Stranded RNA Evolution Fibroblasts Frameshift Mutation Gene Amplification Genes Genetic Genetic Transcription Health Host Defense Human Immune response Immunization Immunoblotting Immunofluorescence Immunologic Immunoprecipitation Infection Isatin Label Macaca mulatta Mass Spectrum Analysis Medical Messenger RNA Metabolic Pathway Missense Mutation Modeling Molecular Monitor Mutation Oncolytic Pathway interactions Pharmaceutical Preparations Phosphorylation Poxviridae Production Proteins RNA RNA Binding Race Regulation Resistance Ribonucleases Risk Role Serial Passage Series Smallpox Viruses System Technology Testing Thiosemicarbazones Transcript Tropism Vaccinia Vaccinia virus Vaccinium Variant Viral Viral Proteins Virus Virus Replication arm base cell type clinical application deep sequencing gene function improved insight mutant oncolytic vector pathogen protein expression protein kinase R protein phosphatase inhibitor-2 public health relevance research study transmission process vector vaccine viral RNA

项目摘要

DESCRIPTION (provided by applicant): In addition to their medical importance as pathogens, vaccine vectors and oncolytic agents, poxviruses offer tractable model systems for discovering fundamental molecular and evolutionary mechanisms underlying the "arms races" between viruses and the host cells they parasitize. These conflicts drive rapid evolution of host and viral proteins and can create species barriers to viral transmission. One such case is the dsRNA-activated protein kinase R (PKR) pathway, which has broad antiviral activity but can be blocked, in a species-specific manner, by a variety of viral factors. Experimental evolution coupled with deep sequencing technology offers a powerful strategy for uncovering new factors and mechanisms underlying the arms race between PKR and its antagonists. For example, serial passage of a vaccinia virus (VV) mutant that encodes an antagonist that only weakly blocks PKR in one cell type yielded progeny viruses that were able to replicate much more efficiently not only in the cells used for the serial passage but also in cells from other species. Further studies revealed that amplification of the gene encoding the weak antagonist or mutations in either of two other VV genes, A24R and A35R, neither of which has previously been implicated as a modulator of the PKR pathway, dramatically improves VV replication in cells from multiple species. The proposed studies aim to elucidate the mechanisms by which mutations in A24R and A35R enable VV overcome the PKR and related host defense systems. Thus, this project will reveal new insights into the functions of these conserved poxvirus genes and into the roles and regulation of the PKR pathway and its VV antagonists. More generally, the results will aid in understanding how large DNA viruses can evolve to overcome host defenses and will have implications for understanding the risks and barriers to cross species transmission of viruses.

描述（由申请人提供）：除了作为病原体、疫苗载体和溶瘤剂的医学重要性外，痘病毒还提供易于处理的模型系统，用于发现病毒与其寄生的宿主细胞之间“军备竞赛”的基本分子和进化机制。这些冲突推动了宿主和病毒的快速进化蛋白质，可以对病毒传播形成物种屏障。其中一个例子是 dsRNA 激活蛋白激酶 R (PKR) 通路，该通路具有广泛的抗病毒活性，但可以通过多种病毒因子以物种特异性方式阻断。实验进化与深度测序技术相结合，为揭示 PKR 及其拮抗剂之间的军备竞赛背后的新因素和机制提供了强大的策略。例如，痘苗病毒（VV）突变体编码的拮抗剂仅在一种细胞类型中微弱地阻断 PKR，其连续传代产生的子代病毒不仅能够在用于连续传代的细胞中更有效地复制，而且能够在来自其他物种的细胞。进一。拟议的研究旨在阐明 A24R 和 A35R 突变使 VV 克服 PKR 和相关宿主防御系统的机制。因此，该项目将揭示对这些保守痘病毒基因的功能以及 PKR 途径及其 VV 拮抗剂的作用和调节的新见解。更一般地说，这些结果将有助于了解大型 DNA 病毒如何进化以克服宿主防御，并将对了解病毒跨物种传播的风险和障碍产生影响。