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：
8848032
负责人：
ADAM P. GEBALLE
金额：
$ 26.4万
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 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8848032
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 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 adaptive immunity 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 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）：除了病原体，疫苗向量和溶瘤剂的医学重要性外，蛇毒还提供了可拖动的模型系统，用于发现病毒和宿主细胞之间“武器种族”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发展蛋白质，可以创建病毒传播的物种障碍。一种情况是DSRNA激活的蛋白激酶R（PKR）途径，该途径具有广泛的抗病毒活性，但可以通过多种病毒因素以物种特异性的方式阻止。实验进化加上深层测序技术，为发现PKR及其对手之间的军备竞赛的新因素和机制提供了有力的策略。例如，编码一个拮抗剂的离子病毒（VV）突变体的连续通过，该拮抗剂仅弱阻止一种细胞类型的PKR产生的后代病毒，这些病毒能够复制得更有效地不仅在用于串行传递的细胞中，而且在细胞中也来自其他物种。进一步的研究表明，在其他两个VV基因A24R和A35R中，编码弱拮抗剂或突变的基因的扩增，以前均未牵涉到PKR途径的调节剂中，大大改善了来自多个物种的细胞中VV复制。拟议的研究旨在阐明A24R和A35R中突变使VV能够克服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的痘病毒基因功能以及PKR途径及其VV拮抗剂的作用和调节的新见解。更一般而言，结果将有助于了解大型DNA病毒可以演变以克服宿主防御能力，并将对了解跨物种传播病毒的风险和障碍具有影响。