Interferon Gene Expression in VSV-Infected Cells

VSV 感染细胞中的干扰素基因表达

• 批准号: 6754765
• 负责人: MAUREEN C FERRAN
• 金额: $ 20.67万
• 依托单位: ROCHESTER INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6754765
• 关键词: I kappa B beta; Vesiculovirus; enzyme activity; gene expression; gene induction /repression; genetic transcription; host organism interaction; immunogenetics; interferons; microorganism immunology; molecular cloning; nuclear factor kappa beta; nucleic acid sequence; phosphorylation; polymerase chain reaction; protein degradation; transfection /expression vector; virus genetics; virus infection mechanism; virus protein

Viral infections commonly trigger the transcription of antiviral genes, including type 1 interferon (IFN). Newly produced IFNs are released from the cell, and activate a signal transduction cascade that induces the transcription of many interferon responsive genes whose products set up an antiviral state in uninfected neighboring cells. Many viruses have evolved to counteract this host defense mechanism, allowing virus replication to occur. The overall focus of this research is to investigate the balance between the ability of the host to induce the antiviral IFN response, and the ability of vesicular stomatitis virus (VSV) to block induction of this system. The cytotoxic VSV matrix (M) protein inhibits host-directed transcription and limits IFN gene expression in VSV-infected cells. The goal of this project is to determine if M protein is solely responsible for suppression of IFN gene expression based on its ability to inhibit host transcription, or if there is a specific mechanism by which VSV regulates IFN gene expression. Our preliminary data suggest that the later is true. We've demonstrated that NF-kappaB activation, an upstream activator of IFN gene expression, is regulated differently in cells infected with an IFN-inducing and IFN-suppressing strains of VSV. We propose that a second viral function inhibits NF-kappaB activation. This viral component would therefore play a crucial role in induction of IFN gene expression. To test this hypothesis, the specific aims to be addressed are: (1) To characterize the VSV Indiana Hr genes from an IFN-suppressing and IFN-inducing strain of VSV by cloning and determining the sequence of each viral gene. (2) To identify the viral component responsible for activation of NF-kappaB, we will investigate NF-kappaB activation in cells infected with recombinant M-defective strains of VSV, as well as determining if activation of NF-kappaB is suppressed in cells transfected with an expression vector encoding one of the viral genes. (3) To understand the mechanism by which the virus regulates NF-kappaB activation, we will determine if IkappaB has been degraded and investigate the phosphorylation status of the IkappaB Kinase in VSV-infected and transfected cells. Upon completion of this project, we will gain new insights into virus modulation of the host interferon response. Detailed knowledge of this cellular response to viral infection could be useful to the development of new antiviral and anticancer therapies.

病毒感染通常会触发抗病毒基因的转录，包括 1 型干扰素 (IFN)。新产生的干扰素从细胞中释放出来，并激活信号转导级联，诱导许多干扰素反应基因的转录，这些基因的产物在未感染的邻近细胞中建立抗病毒状态。许多病毒已经进化到抵消这种宿主防御机制，从而允许病毒复制。本研究的总体重点是研究宿主诱导抗病毒 IFN 反应的能力与水泡性口炎病毒 (VSV) 阻断该系统诱导的能力之间的平衡。细胞毒性 VSV 基质 (M) 蛋白抑制宿主定向转录并限制 IFN 基因 VSV 感染细胞中的表达。该项目的目标是确定 M 蛋白是否仅根据其抑制宿主转录的能力负责抑制 IFN 基因表达，或者是否存在 VSV 调节 IFN 基因表达的特定机制。我们的初步数据表明后者是正确的。我们已经证明，NF-kappaB 激活（IFN 基因表达的上游激活剂）在感染 IFN 诱导型和 IFN 抑制型 VSV 菌株的细胞中受到不同的调节。我们提出第二种病毒功能抑制 NF-kappaB 激活。因此，这种病毒成分将发挥至关重要的作用 诱导 IFN 基因表达。为了检验这一假设，需要解决的具体目标是：（1）通过克隆和确定每个病毒基因的序列来表征来自 IFN 抑制和 IFN 诱导 VSV 菌株的 VSV Indiana Hr 基因。 (2) 为了鉴定负责 NF-kappaB 激活的病毒成分，我们将研究被重组 M 缺陷型 VSV 菌株感染的细胞中 NF-kappaB 的激活，并确定转染细胞中 NF-kappaB 的激活是否受到抑制具有编码病毒基因之一的表达载体。 (3) 为了了解病毒调节NF-kappaB激活的机制，我们将确定IkappaB是否已被降解并研究其 VSV 感染和转染细胞中 IkappaB 激酶的磷酸化状态。该项目完成后，我们将获得关于病毒调节宿主干扰素反应的新见解。对这种细胞对病毒感染的反应的详细了解可能有助于开发新的抗病毒和抗癌疗法。

项目成果

The Propagation, Quantification, and Storage of Vesicular Stomatitis Virus.

• DOI: 10.1002/cpmc.110
• 发表时间: 2020-09-01
• 期刊: Current protocols in microbiology
• 作者: Abdelmageed, Alaa A;Ferran, Maureen C
• 通讯作者: Ferran, Maureen C

Preface. Cardiomyocytes.

前言。

• DOI: 10.1007/978-1-4939-2572-8
• 发表时间: 2015
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Skuse,GaryR;Ferran,MaureenC
• 通讯作者: Ferran,MaureenC

The vesicular stomatitis virus matrix protein inhibits NF-κB activation in mouse L929 cells.

• DOI: 10.1016/j.virol.2016.09.009
• 发表时间: 2016-12
• 期刊: VIROLOGY
• 影响因子: 3.7
• 作者: Varble, Andrew J.;Ried, Christopher D.;Hammond, Warren J.;Marquis, Kaitlin A.;Woodruff, Matthew C.;Ferran, Maureen C.
• 通讯作者: Ferran, Maureen C.