New Opportunities - Mechanisms of Early Vaccinia Viral Morphogensis

摘要 痘病毒家族的成员已经被研究用于它们作为疫苗和表达载体的应用，并且最近由于它们作为潜在的生物武器的用途而被专注地研究。痘苗病毒是病毒的原型成员，通过多种形式进化获得感染性，早期膜形态发生在其中起着关键作用。即使病毒通常篡夺宿主因子的使用，它仍然不清楚是否是众所周知的细胞室的膜形态发生中起作用的关键宿主转运因子参与早期牛痘膜形态发生。我们现在已经发现了coatomer的作用，coatomer是一种宿主蛋白质复合物，已知可形成Coat Protein I（COP！）运输囊泡然而，对外壳体如何参与早期病毒形态发生的深入研究表明，病毒绕过了形成宿主COP 1囊泡的关键调控机制，而是与两个 蛋白质、病毒K7和宿主KDEL受体（KDELR）通过关键的二赖氨酸残基的相互作用可能是重要的。因此，为了进一步了解外壳体及其两种相互作用的蛋白质如何促进病毒复制，我们提出了两个主要目标。在由维克托许（Victor Hsu）领导的一个目标中，NERCE提议资助，将检查外被体的扰动在潜在积累特定病毒形式中的作用。此外，将检查KDELR在病毒形式上的分布，以及是否 其配体在病毒复制中起作用。作为Stuart lsaacs领导的另一个目标，在MARCE的资助下，将检查病毒K7蛋白在其缺失时的影响，以及当表达不能与外被体相互作用的突变形式时的影响。 这一合作努力以多种方式回应了两个区域英才中心根据“新机会”倡议提出的申请。首先，我们将深入了解A类病原体，我们不仅将阐明宿主蛋白在病毒复制过程中发挥作用的新机制，而且还将确定未来设计的潜在关键靶标。 针对病毒最丰富的传染形式进行干预。第二，该提案的资金将允许主要研究者之一（维克托许）与痘病毒专家（斯图尔特艾萨克斯）合作，从而将囊泡运输机制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 除了完全新颖的技术外，这项合作的互补性专业知识可能会加强对牛痘病毒如何与其宿主相互作用的阐明。第三，该提案的潜在资金可能会为未来最终更全面的应用奠定基础，例如通过NIH R 01机制。

Abstract 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 acquiring infectivity, for which early 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 cellular compartments participate in early vaccinia membrane morphogenesis. We have now discovered a role for coatomer, a host protein complex known to shape Coat Protein I (COP!) transport vesicles. However, insights into how coatomer participates in early viral morphogenesis reveals that the virus bypasses key regulatory mechanisms that form host COP1 vesicles, but instead its interaction with two proteins, the viral K7 and the host KDEL receptor (KDELR) through critical di-lysine residues is likely important. 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 lsaacs 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.