Cellular interactions of viral matrix protein

病毒基质蛋白的细胞相互作用

• 批准号: 6616565
• 负责人: DOUGLAS S. LYLES
• 金额: $ 28.8万
• 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 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6616565
• 关键词: DNA directed RNA polymerase; HeLa cells; SDS polyacrylamide gel electrophoresis; Vesiculovirus; apoptosis; gene expression; genetic transcription; interferons; mass spectrometry; microarray technology; permease; polymerase chain reaction; tissue /cell culture; transcription factor; virus infection mechanism; virus protein; western blottings

DESCRIPTION (provided by applicant): Many viruses inhibit the expression of host genes. In most cases the role of the virus-induced inhibition of host gene expression is to inhibit the host antiviral response, particularly the synthesis of type I (alpha/beta) interferons. An important consequence of the inhibition of host gene expression is that it may promote programmed cell death or apoptosis in host cells. The proposed experiments address the mechanisms of inhibition of host gene expression and the consequences of this inhibition for viral induction of an interferon response or induction of apoptosis using the prototype rhabdovirus, vesicular stomatitis virus (VSV). We have established that the viral matrix (M) protein plays a major role in the inhibition of host gene expression by VSV. Aim 1 is to determine the mechanism of inhibition of host transcription by M protein. We have shown that inhibition of host RNA polymerase II-dependent transcription is due to inactivation of the basal transcription factor TFIID. We will determine changes that have occurred in TFIID from infected cells that account for the inhibition using immuno-affinity purified, epitope-tagged TFIID. Aim 2 is to determine the mechanism by which M protein regulates type I interferon gene expression in host cells. We will determine the extent to which M protein inhibits interferon gene expression by inhibition of basal transcription factors, upstream activators of interferon gene expression, or nuclear-cytoplasmic transport of interferon mRNA. Aim 3 is to determine the mechanism by which M protein regulates induction of apoptosis in host cells. We have established that M protein is a potent inducer of apoptosis when expressed in the absence of other viral components. However, there are at least two VSV components that contribute to induction of apoptosis in the context of a virus infection. We will analyze the molecular components involved in activation of apoptosis by M protein and the other viral component. Infectious VSV cDNA clones will be used to identify other viral components involved in the induction of apoptosis by mapping the sequences of previously isolated viral mutants that enhance their ability to induce apoptosis. These experiments will provide a clearer understanding of the molecular basis for viral pathogenesis including the mechanism for the inhibition of host transcription, the inhibition of interferon production, and the activation of apoptosis.

描述（由申请人提供）：许多病毒抑制宿主基因的表达。在大多数情况下，病毒诱导的宿主基因表达抑制的作用是抑制宿主抗病毒反应，尤其是I型（alpha/beta）干扰素的合成。抑制宿主基因表达的重要结果是，它可能促进宿主细胞中编程的细胞死亡或凋亡。该提出的实验解决了抑制宿主基因表达的机制，以及这种抑制这种抑制干扰素反应的抑制作用或使用原型的色齿病毒（VSICULAL OTAMATIL PIRUS）（VSV）诱导了干扰素反应或诱导凋亡的后果。我们已经确定，病毒基质（M）蛋白在VSV抑制宿主基因表达中起主要作用。目的1是确定M蛋白抑制宿主转录的机制。我们已经表明，抑制宿主RNA聚合酶II依赖性转录是由于基础转录因子TFIID的失活所致。我们将确定感染细胞中TFIID中发生的变化，这些细胞使用免疫亲和力纯化的表位标记的TFIID来解释抑制作用。目标2是确定M蛋白调节宿主细胞中I型干扰素基因表达的机制。我们将通过抑制基础转录因子，干扰素基因表达的上游激活剂或干扰素mRNA的核胞质转运来确定M蛋白在多大程度上抑制干扰素基因的表达。目标3是确定M蛋白调节宿主细胞凋亡诱导的机制。我们已经确定，在没有其他病毒成分的情况下表达时，M蛋白是凋亡的有效诱导剂。但是，在病毒感染的情况下，至少有两个VSV成分有助于诱导凋亡。我们将分析M蛋白和其他病毒成分激活凋亡的分子成分。传染性VSV cDNA克隆将通过映射先前分离的病毒突变体的序列来鉴定参与诱导凋亡的其他病毒成分，从而增强其诱导凋亡的能力。这些实验将对病毒发病机理的分子基础有更清晰的了解，包括抑制宿主转录的机制，抑制干扰素产生和凋亡的激活。