Mechanisms of vaccinia virus dissemination

痘苗病毒传播机制

基本信息

批准号：
8662699
负责人：
HERVE F AGAISSE
金额：
$ 20.81万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-16 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8662699
关键词：
Actins Adaptor Signaling Protein Address Boxing Cell membrane Cell physiology Cells Complex Cytoskeleton Disease Outbreaks Docking EPHA8 gene Environment Event Family Family member Foundations Genetic Goals Image Infection Intracellular Membranes Investigation Knowledge Life Cycle Stages Light Link Mediating Membrane Membrane Proteins Molecular Monkeypox Orthopoxvirus Pathogenesis Phosphorylation Phosphotransferases Poxviridae Preventive Intervention Production Protein Tyrosine Kinase Proteins RNA Interference Receptor Protein-Tyrosine Kinases Recruitment Activity Research Role Satellite Viruses Seminal Signal Transduction Site Smallpox Smallpox Viruses Tail Testing Therapeutic Intervention Tyrosine Vaccines Vaccinia Vaccinia virus Vaccinium Viral Viral Proteins Virus base cell motility cellular imaging design disorder prevention driving force epizootic extracellular insight microbial pathogen polymerization programs public health relevance receptor src-Family Kinases trans-Golgi Network transmission process virus genetics

项目摘要

DESCRIPTION (provided by applicant): The Orthopoxvirus genus of the family Poxviridae includes both the causative agent of smallpox, variola virus, and the vaccine strain used for smallpox eradication, Vaccinia virus. Even though naturally occurring smallpox has been successfully eradicated, we need to better understand the mechanisms supporting Orthopoxviruses dissemination in light of potential bioterrorist attacks and/or outbreaks of epizootic infections (monkeypox). The life cycle of Orthopoxviruses relies on the production of two infectious forms, the intracellular mature viruses (IMV) and the extracellular viruses (EV). EV is formed through wrapping of IMV in double membranes derived from the trans-Golgi network. The fusion of the outer membrane of EV to the plasma membrane leads to formation of cellular-associated EV (CEV). A central event in CEV dissemination is phosphorylation of the viral protein A36 by non-receptor tyrosine kinases of the Src family. A36 phosphorylation on tyrosine residues creates docking sites for the adaptor proteins Nck1 and Grb2. These adaptors mediate the recruitment of N- WASP, which mediates the recruitment and activation of the actin nucleator ARP2/3. In contrast to the molecular machinery leading to actin tail formation, the identity of the putative receptor tyrosine kinase mediating Src activation is unknown. Moreover, the identity of the viral protein that may engage this putative receptor is unknown as well. To address this gap in knowledge, we have combined our extensive expertise in the mechanisms supporting actin-based motility, high-throughput imaging and large-scale RNAi-based genetic investigation. In this R21 exploratory application, we propose (Aim1) to determine the receptor tyrosine kinase(s) involved in actin tail formation and (Aim2) to determine the viral protein(s) engaging receptor tyrosine kinase signaling at the plasma membrane. These exploratory studies may provide mechanistic insight into molecular and cellular processes relevant to the treatment and prevention of diseases caused by pathogens that exploit the actin cytoskeleton for their dissemination.

描述（由申请人提供）：家族鼠科的正托病毒属包括天花，Variola病毒的病原体，以及用于消除天花的疫苗菌株，即节日病毒。即使自然存在的天花已经成功地消除了，我们仍需要更好地了解支持正托病毒传播的机制，鉴于潜在的生物恐怖袭击和/或epizootic感染的爆发（Monkeypox）。正托病毒的生命周期依赖于两种传染性形式的产生，细胞内成熟病毒（IMV）和细胞外病毒（EV）。 EV是通过从反式高尔基网络得出的双膜中包裹的IMV形成的。 EV向质膜的外膜融合导致细胞相关EV（CEV）的形成。 CEV传播中的一个中心事件是SRC家族的非受体酪氨酸激酶对病毒蛋白A36的磷酸化。酪氨酸残基上的A36磷酸化为衔接蛋白NCK1和GRB2创建对接位点。这些衔接子介导了N-WASP的募集，该衔接介导了肌动蛋白成核ARP2/3的募集和激活。与导致肌动蛋白尾部形成的分子机械相反，推定的受体酪氨酸激酶介导SRC激活的身份尚不清楚。此外，可能吸引这种推定受体的病毒蛋白的身份也未知。为了解决知识的这一差距，我们将广泛的专业知识结合在支持基于肌动蛋白的运动，高通量成像和大规模RNAi基因遗传研究的机制中。在此R21探索性应用中，我们建议（AIM1）确定参与肌动蛋白尾部形成的受体酪氨酸激酶，并（AIM2）确定质膜上参与受体酪氨酸激酶信号的病毒蛋白。这些探索性研究可能会提供有关与病原体引起的治疗和预防疾病相关的分子和细胞过程的机械洞察力，这些病原体利用了肌动蛋白细胞骨架的传播。