CELLULAR INTERACTIONS OF VIRAL MATRIX PROTEIN

病毒基质蛋白的细胞相互作用

• 批准号: 6149778
• 负责人: DOUGLAS S. LYLES
• 金额: $ 28.24万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6149778
• 关键词: DNA binding protein; DNA directed RNA polymerase; Vesiculovirus; genetic transcription; intracellular transport; tissue /cell culture; virus cytopathogenic effect; virus infection mechanism; virus protein

The mechanisms by which viruses cause pathological changes in the cells they infect is an area of active current interest. In the case of vesicular stomatitis virus (VSV), the prototype rhabdovirus, a viral structural protein, the matrix (M) protein, appears to be responsible for several of the major cytopathic effects of virus infection. M protein has the ability to inhibit host-directed gene expression and to cause the characteristic rounding of VSV-infected cells when expressed in the absence of other viral components. The goal of this project is to determine the mechanism of VSV-induced cytopathic effects and to identify strategies for enhancing the resistance of host cells to the cytopathic effects of virus infection. The experiments proposed here are focused primarily on the mechanism by which VSV inhibits host- directed transcription and involve the following Specific Aims: Aim 1 is to determine the mechanism of inhibition of transcription initiation by nuclear extracts from VSV-infected cells. Our recent data indicate that virus-induced inhibition of transcription by host RNA polymerase II involves defective activity of basal transcription initiation factors and that recombinant TATA-binding protein (TBP) subunit of transcription factor IID (TFIID) can overcome the defect in nuclear extracts from VSV- infected cells. How TBP in inactivated will be determined by analysis of TBP expression and activity of TFIID purified from nuclear extracts from VSV-infected cells. We will also determine whether overexpression of TBP in vivo make cells resistant to the inhibition of host transcription by VSV. Aim 2 is to determine whether viral components other than M protein are involved in the inhibition of transcription in VSV-infected cells. This hypothesis will be tested by generating a VSV deletion mutant lacking an M gene and testing its effects on host gene expression. Upon completion of these experiments we will have identified molecular targets involved in the inhibition of host transcription by VSV, and we will have a more complete view of the viral components involved in the inhibition. These experiments should also point the way to strategies for making cells resistant to the cytopathic effects of virus infection, either by overexpressing transcription factor(s) involved in the inhibition or by genetically engineering transcription factors that are resistant to the inhibitory effects of virus infection. Cells that are resistant to one or more cytopathic effects of virus infection will constitute a valuable tool in future studies of viral pathogenesis and may also be of practical value in developing antiviral strategies in the intact host.

病毒引起细胞病理变化的机制 它们感染的是当前人们感兴趣的一个领域。 如果是 水泡性口炎病毒（VSV），原型弹状病毒，一种病毒 结构蛋白，即基质 (M) 蛋白，似乎是造成这种情况的原因 对于病毒感染的几种主要细胞病变效应。 中号 蛋白质具有抑制宿主定向基因表达的能力 表达时导致 VSV 感染细胞特征性变圆 在没有其他病毒成分的情况下。 该项目的目标是 确定 VSV 诱导的细胞病变效应的机制并 确定增强宿主细胞抵抗力的策略 病毒感染的细胞病变效应。 这里提出的实验 主要关注 VSV 抑制宿主的机制 定向转录并涉及以下具体目标：目标 1 目的是确定抑制转录起始的机制 通过 VSV 感染细胞的核提取物。 我们最近的数据表明 病毒诱导宿主 RNA 聚合酶抑制转录 II涉及基础转录起始因子的活性缺陷 以及重组 TATA 结合蛋白 (TBP) 转录亚基 IID因子（TFIID）可以克服VSV-核提取物的缺陷 被感染的细胞。 通过分析确定TBP如何灭活 从核提取物中纯化的 TFIID 的 TBP 表达和活性 来自 VSV 感染的细胞。 我们还将确定是否过度表达 TBP在体内使细胞抵抗宿主的抑制 通过VSV转录。 目标 2 是确定病毒成分是否 除 M 蛋白外，其他蛋白也参与转录抑制 VSV 感染的细胞。 该假设将通过生成 VSV 进行检验 缺乏M基因的缺失突变体并测试其对宿主基因的影响 表达。 完成这些实验后我们将得到 确定了参与抑制宿主的分子靶标 通过 VSV 转录，我们将对病毒有更全面的了解 参与抑制的成分。 这些实验还应该 为使细胞抵抗细胞病变的策略指明道路 病毒感染的影响，或者通过过度表达转录 参与抑制或基因工程的因素 转录因子对抑制作用有抵抗力 病毒感染。对一种或多种细胞病变有抵抗力的细胞 病毒感染的影响将成为未来的一个有价值的工具 病毒发病机制的研究，也可能具有实用价值 在完整宿主中制定抗病毒策略。