The Role of Cellular Chaperones in RNA Virus Infection

细胞伴侣在 RNA 病毒感染中的作用

• 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
• 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

 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（由申请人提供）：一个功能蛋白质稳态网络，包括细胞伴侣和泛素-蛋白酶体系统，对细胞活力至关重要 和人类健康。越来越多的与蛋白质折叠功能障碍相关的疾病证明了这一点。病毒感染还对细胞蛋白质稳态机制造成重大负担，这是由于感染期间存在大量的少数结构复杂的蛋白质。我们以前已经表明，90 kDa的热休克蛋白，热休克蛋白90的化学抑制剂，通过抑制其衣壳前体蛋白，P1的加工，有效地阻止许多肠道病毒物种的感染。值得注意的是，在所有测试的病毒中，即使在长期传代后，也没有出现耐药病毒变体，这表明导致P1折叠的突变不依赖于Hsp 90降低了病毒的适应性。这得到了以下观察结果的支持：选择对选择后持续存在的病毒群体的组成具有显著影响。该提案的目标是（i）利用新的深度测序技术和群体遗传学来量化Hsp 90抑制剂对病毒群体结构的影响，以及（ii）对选定群体中观察到的伴侣蛋白独立性机制进行生物药理学表征。我们的工作假设是Hsp 90功能的抑制将改变病毒群体结构和病毒基因组序列，以反映对病毒蛋白质折叠的改变的约束。