The Role of Cellular Chaperones in RNA Virus Infection

细胞伴侣在 RNA 病毒感染中的作用

基本信息

批准号：
9197311
负责人：
Patrick Timothy Dolan
金额：
$ 5.71万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9197311
关键词：
Address Antiviral Agents Antiviral Therapy Biochemical Biological Assay Biophysics Capsid Chemicals Client Dependence Development Disease Enterovirus Evolution Exhibits Experimental Designs Family Picornaviridae Functional disorder Future Genetic Goals HSP 90 inhibition Health Heat shock proteins Homeostasis Human Human poliovirus Infection Kinetics Laboratories Lead Molecular Chaperones Monitor Mutation Pathway interactions Population Population Dynamics Population Genetics Positioning Attribute Process Protein Precursors Proteins Proteolytic Processing Quality Control RNA Virus Infections Resistance Resolution Role Structure System Technology Testing Translations Ubiquitin Variant Viral Viral Genome Viral Pathogenesis Viral Proteins Virus Virus Diseases Work biophysical techniques clinically relevant cofactor deep sequencing experimental study fitness functional genomics inhibitor/antagonist innovation interest multicatalytic endopeptidase complex novel protein complex protein folding protein function proteostasis public health relevance ribosome profiling screening viral fitness

项目摘要

DESCRIPTION (provided by applicant): A functioning protein homeostasis network, which includes cellular chaperones and the ubiquitin-proteasome system, is critical to cellular viability and human health. This is evidenced by an ever-growing number of diseases associated with protein folding dysfunction. Viral infection also places a significant burden on the cellular proten homeostasis machinery, due to the large amount of a few structurally complex proteins during infection. We have shown previously that chemical inhibitors of the 90kDa heat-shock protein, Hsp90, potently block infection by numerous enteroviral species by inhibiting the processing of their capsid precursor protein, P1. Notably, in all of the viruses tested, resistant viral variantsdid not emerge, even after long-term passage suggesting that mutations that lead to folding of P1 independent of Hsp90 reduce fitness of the virus. This is supported by the observation that selection has significant effects on the composition of the viral population that persists after selection. The goal of this proposal is (i) to leverage new deep-sequencing technologies and population genetics to quantify the effect of Hsp90 inhibitors on viral population structure, and (ii) to biophysically characterize the mechanism of chaperone independence observed in the selected population. Our working hypothesis is that the inhibition of Hsp90 function will change the viral population structure and viral genome sequence to reflect the altered constraints on viral protein folding.

描述（应用程序提供）：功能性蛋白质稳态网络，包括细胞伴侣和泛素 - 蛋白酶体系统，对细胞生存能力至关重要和人类健康。与蛋白质折叠功能障碍相关的疾病数量不断增长，这证明了这一点。病毒感染还对细胞蛋白稳态机制进行了重大燃烧，这是由于感染过程中有大量结构复杂的蛋白质。我们先前已经证明，90KDA热休克蛋白HSP90的化学抑制剂可能通过抑制其Capsid前体蛋白P1的加工来阻断许多肠病毒物种的感染。值得注意的是，在所有测试的病毒中，即使长期通过后，也不会出现耐药性病毒变异剂，这表明导致P1折叠的突变独立于HSP90降低了病毒的适应性。这是由观察结果支持的，即选择会对选择后持续存在的病毒种群的组成具有重大影响。该提案的目的是（i）利用新的深层测序技术和种群遗传学来量化HSP90抑制剂对病毒种群结构的影响，（ii）以生物物理表征所选人群中观察到的链酮独立性的机制。我们的工作假设是，HSP90功能的抑制作用将改变病毒种群结构和病毒基因组序列，以反映病毒蛋白折叠的约束改变。