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

痘病毒家族的成员因其作为疫苗和表达的应用而受到调查 最近，由于它们被用作潜在的生物武器，人们对它们进行了认真的研究。牛痘 病毒的原型成员通过多种形式进化，其中膜形态发生起关键作用 角色。尽管病毒通常会篡夺宿主因素来使用它，但目前尚不清楚关键宿主是否 已知在宿主细胞室的膜形态发生中起作用的运输因子 参与早期牛痘疫苗膜形态的形成，这是获得病毒感染性的关键过程。我们 现在已经确定了辅酶分子的作用，这是一种宿主蛋白复合体，其特征是形成外壳蛋白I (Copi)运输小泡。然而，对它如何参与早期病毒形态发生的洞察揭示了 病毒绕过了形成宿主COPI囊泡的关键调控机制，而是与 两种蛋白质，病毒K7和宿主KDEL受体(KDELR)通过关键的二赖氨酸残基在这些 相互作用的蛋白质。因此，为了进一步了解辅酶及其两个相互作用的蛋白质是如何 为了促进病毒复制，我们提出了两个主要目标。在由Victor Hsu领导的一个目标中，提出了 来自NERCE的资金，将检查辅助器的扰动在潜在积累中的影响 特殊的病毒形式。此外，将检查KDELR在病毒形式上的分布情况 以及它的配体是否在病毒复制中发挥作用。作为斯图尔特·艾萨克领导的另一个目标 由Marce提议的资助，将检查病毒K7蛋白在其缺失时的影响，还 当表达一种不能与辅酶原相互作用的突变形式时。 这一协作努力以多种方式响应了根据《新的 由两个区域英才中心提出的“机遇”倡议。首先，我们将获得 对A类病原体的洞察，我们不仅将阐明宿主通过哪些新机制 蛋白质在病毒复制过程中的功能，也为未来设计确定潜在的关键靶点(S) 对病毒最丰富的传染性形式的干预。第二，这项提案的资金将 允许主要调查人员之一(Victor Hsu)与痘病毒专家(Stuart Isaacs)合作，以及 因此，将泡运输机制方面的外部专业知识应用于痘病毒生物学中的关键问题。 因为病毒通常征用宿主机制来与宿主交互，而不是发明 完全是新的，合作的互补专业知识可能会增强对 牛痘病毒如何与宿主相互作用。第三，这项提案的潜在资金可能会为 最终在未来得到更全面的应用，例如通过NIH R01机制。

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.