New Ops: Mechanisms of early vaccinia viral morphogenesis (trans-RCE proj)

新操作：早期痘苗病毒形态发生的机制（trans-RCE proj）

• 批准号: 7645453
• 负责人: VICTOR W HSU
• 金额: $ 24.44万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7645453
• 关键词: Biological; Biology; Bypass; Capsid Proteins; Categories; Coat Protein Complex I; Coatomer Protein; Collaborations; Complex; Family; Funding; Future; Golgi Apparatus; Head; Hospitals; Integration Host Factors; KDEL receptor; Ligands; Membrane; Morphogenesis; Pennsylvania; Play; Poxviridae; Process; Proteins; Request for Applications; Research Personnel; Research Project Grants; Role; Staging; Transport Vesicles; United States National Institutes of Health; Universities; Vaccines; Vaccinia; Vaccinia virus; Vesicle; Viral; Viral Proteins; Virus; Woman; expression vector; insight; member; mutant; novel; pathogen; protein function; therapy design

Members of the poxvirus family have been investigated for their applications as vaccines and expression vectors, and more recently intently studied because of their use as potential biological weapons. Vaccinia virus, the prototypic member, evolves through multiple forms in which membrane morphogenesis plays a key role. Even though viruses generally usurp host factors for its use, it remains unclear whether key host transport factors that are well known to act in membrane morphogenesis of host cellular compartment participate in early vaccinia membrane morphogenesis, a process critical in acquiring viral infectivity. We have now identified a role for coatomer, a host protein complex well characterized to form Coat Protein I (COPI) transport vesicles. However, insights into how it participates in early viral morphogenesis reveals that the virus bypasses key regulatory mechanisms that form host COPI vesicles, but instead interacts with two proteins, the viral K7 and the host KDEL receptor (KDELR) through critical di-Iysine residues on these interacting proteins. Thus, to gain further insight into how coatomer and its two interacting proteins contribute to viral replication, we propose two major aims. In one aim headed by Victor Hsu with proposed funding from NERCE, perturbation of coatomer will be examined for its effect in potentially accumulating particular viral forms. Moreover, the KDELR will be examined with respect to its distribution on viral forms and also whether its ligands have a role in viral replication. As another aim headed by Stuart Isaacs with proposed funding from MARCE, the viral K7 protein will be examine for effects upon its deletion and also when a mutant form that cannot interact with coatomer is expressed. This collaborative effort is responsive in multiple ways to the request for application under the "New Opportunities" initiative by the two respective Regional Centers of Excellence. First, we will be gaining insights into a Category A pathogen, for which we will not only elucidate novel mechanisms by which a host protein functions dUring viral replication, but also identify potential key target(s) for the future design of intervention against the most abundant infectious form of the virus. Second, funding of this proposal will allow one of the main investigators (Victor Hsu) to collaborate with a poxvirus expert (Stuart Isaacs), and thus, applying outside expertise in mechanisms of vesicular transport to critical issues in poxvirus biology. As viruses generally commandeer host mechanisms for their interaction with the host rather than inventing completely novel ones, the complementary expertise of the collaboration will likely enhance the elucidation of how vaccinia virus interacts with its host. Third, potential funding of this proposal will likely set the stage for an eventual more comprehensive application in the future, such as through the NIH R01 mechanism.

痘病毒家族成员已被研究作为疫苗和表达的应用 载体，并且最近因其用作潜在的生物武器而被深入研究。痘苗病毒 病毒作为原型成员，通过多种形式进化，其中膜形态发生起着关键作用 角色。尽管病毒通常会篡夺宿主因子以供其使用，但仍不清楚关键宿主是否 众所周知在宿主细胞区室膜形态发生中起作用的转运因子 参与早期痘苗膜形态发生，这是获得病毒感染性的关键过程。我们 现在已经确定了外壳蛋白的作用，外壳蛋白是一种宿主蛋白​​复合物，经过充分表征，可形成外壳蛋白 I (COPI) 运输囊泡。然而，对其如何参与早期病毒形态发生的深入了解揭示了 该病毒绕过了形成宿主 COPI 囊泡的关键调节机制，而是与 两种蛋白质，即病毒 K7 和宿主 KDEL 受体 (KDELR)，通过这些蛋白质上的关键二赖氨酸残基 相互作用的蛋白质。因此，为了进一步了解涂层异构体及其两种相互作用的蛋白质如何 有助于病毒复制，我们提出两个主要目标。在 Victor Hsu 领导的一项目标中，提出了 在 NERCE 的资助下，将检查涂层异构体的扰动对潜在累积的影响 特定的病毒形式。此外，还将检查 KDELR 在病毒形式上的分布 以及它的配体是否在病毒复制中发挥作用。作为 Stuart Isaacs 领导的另一个目标 拟议的 MARCE 资助，将检查病毒 K7 蛋白对其删除的影响，以及 当表达不能与涂层异构体相互作用的突变形式时。 这种协作努力以多种方式响应“新 机会”倡议由两个各自的区域卓越中心发起。首先，我们将获得 对 A 类病原体的深入了解，我们不仅会阐明宿主的新机制 蛋白质在病毒复制过程中的功能，而且还确定了未来设计的潜在关键目标 针对最丰富的传染性病毒形式的干预措施。其次，本提案的资助将 允许主要调查员之一（Victor Hsu）与痘病毒专家（Stuart Isaacs）合作，以及 因此，将囊泡运输机制方面的外部专业知识应用于痘病毒生物学的关键问题。 由于病毒通常会征用宿主机制来与宿主相互作用，而不是发明宿主机制 完全新颖的，合作的互补专业知识可能会增强对 痘苗病毒如何与其宿主相互作用。第三，该提案的潜在资金可能会为 未来最终会有更全面的应用，例如通过 NIH R01 机制。