Control of translation by VSV

通过 VSV 控制翻译

基本信息

批准号：
7015597
负责人：
DOUGLAS S. LYLES
金额：
$ 28.03万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-02-01 至 2009-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7015597
关键词：
RNA virus SDS polyacrylamide gel electrophoresis Vesiculovirus flow cytometry gel electrophoresis gene expression genetic mapping genetic translation messenger RNA northern blottings phosphoprotein phosphatase phosphorylation protein biosynthesis protein kinase recombinant virus translation factor virus protein western blottings

项目摘要

DESCRIPTION (provided by applicant): The control of translation plays a critical role in the pathogenesis of many viruses. The inhibition of host protein synthesis is important for suppression of the host antiviral response, while the selective translation of viral mRNAs is important for virus propagation in the host. Translational control by vesicular stomatitis virus (VSV), the prototype rhabdovirus, is one of the best studied examples among the nonsegmented negative stranded RNA viruses. Previous experiments have made it clear that VSV uses novel mechanisms to control translation that are distinct from those of other well-studied RNA viruses, such as picornaviruses and influenza viruses. The goal of this project is to determine these mechanisms by which VSV inhibits host protein synthesis and promotes viral protein synthesis. Previous work has shown that the translation factor elF2alpha is inactivated in VSV-infected cells as a result of phosphorylation by protein kinase R. However, we have shown that the cap-binding elF4F complex is also inactivated in VSV-infected cells. Aim 1 is to determine how elF4F is inactivated, and the relative contribution of elF4F versus elF2alpha inactivation in the inhibition of host protein synthesis in VSV-infected cells. Aim 2 is to determine the viral components responsible for inducing the inhibition of host protein synthesis. These studies will use new recombinant viruses to map the genes of VSV mutants that are either more effective or less effective than wild-type VSV in the inhibition of host translation. Aim 3 is to determine the basis of resistance of viral mRNAs to the inhibition of translation. These experiments will use chimeric mRNAs containing viral or host 5' and 3' untranslated regions to test for the presence of cis-acting sequences that enhance translation of viral mRNAs. We will also test recombinant viruses that express these chimeric mRNAs to determine whether mRNA synthesis by the viral transcriptase confers the resistance to the inhibition of translation. Upon completion of these experiments we will have important new information about how viruses suppress the antiviral response in the host, and how they promote expression of their own gene products. The results of these experiments will be novel, because the mechanisms used by VSV differ substantially from those of other well-studied prototype viruses. Finally, it is very likely that there are host mRNAs that use mechanisms similar to those of VSV mRNAs to enhance their translation under conditions such as stress. Thus we expect to gain new insight into regulation of cellular translation under other conditions where the activities of translation initiation factors are inhibited.

描述（由申请人提供）：翻译的控制在许多病毒的发病机理中起关键作用。抑制宿主蛋白合成对于抑制宿主抗病毒反应很重要，而病毒mRNA的选择性翻译对于宿主的病毒传播很重要。囊泡炎性病毒（VSV）的翻译控制，原型的色齿病毒是未分段的阴性滞留RNA病毒中研究的例子之一。先前的实验表明，VSV使用新型机制来控制与其他良好研究的RNA病毒（例如Picornaviruse and Imparza病毒）不同的翻译。该项目的目的是确定VSV抑制宿主蛋白合成并促进病毒蛋白合成的这些机制。先前的工作表明，由于蛋白激酶的磷酸化，VSV感染的细胞中的翻译因子ELF2Alpha被灭活。但是，我们已经表明，在VSV感染的细胞中，Caping结合ELF4F复合物也被灭活。目标1是确定ELF4F是如何灭活的，以及ELF4F与ELF2Alpha失活在抑制VSV感染细胞中宿主蛋白合成中的相对贡献。目标2是确定负责诱导宿主蛋白合成的病毒成分。这些研究将使用新的重组病毒来映射比野生型VSV在抑制宿主翻译方面更有效或更有效的VSV突变体的基因。目的3是确定病毒mRNA对抑制翻译的抗性的基础。这些实验将使用含有病毒或宿主5'和3'未翻译区域的嵌合mRNA测试，以增强病毒mRNA翻译的顺式作用序列。我们还将测试表达这些嵌合mRNA的重组病毒，以确定病毒转录酶合成的mRNA合成是否赋予对翻译抑制的抗性。这些实验完成后，我们将获得有关病毒如何抑制宿主中抗病毒反应的重要新信息，以及它们如何促进自己的基因产物的表达。这些实验的结果将是新颖的，因为VSV所使用的机制与其他经过深入的原型病毒的机制有很大不同。最后，很有可能有一些使用与VSV mRNA相似的机制在压力等条件下增强其翻译的机制。因此，我们期望在抑制翻译起始因子活性的其他条件下对细胞翻译的调节进行新的见解。