New Opportunities - Mechanisms of Early Vaccinia Viral Morphogensis

摘要 痘病毒家族的成员因其作为疫苗和表达载体的应用而被研究，最近由于它们被用作潜在的生物武器而被集中研究。痘苗病毒是痘苗病毒的原型成员，在获得感染力的过程中通过多种形式进化，其中早期膜形态发生起着关键作用。尽管病毒通常会篡夺宿主因子来使用它，但目前还不清楚在细胞隔膜形态发生中起作用的关键宿主运输因子是否参与了痘苗病毒早期膜形态发生。我们现在已经发现了Coom er的作用，这是一种已知形成Coat Protein I(CoP！)的宿主蛋白复合体。运输囊泡。然而，对Cotom如何参与早期病毒形态发生的洞察表明，该病毒绕过了形成宿主COP1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可能是重要的。因此，为了进一步了解辅酶及其两个相互作用的蛋白如何促进病毒复制，我们提出了两个主要目标。在由Victor Hsu领导的一个目标中，建议由NERCE提供资金，将检查辅助器的扰动在潜在积累特定病毒形式方面的影响。此外，将检查KDELR在病毒形式上的分布情况，以及是否 它的配体在病毒复制中发挥作用。作为Stuart lsaacs领导的另一个目标，建议由Marce提供资金，将检查病毒K7蛋白在其缺失时的影响，以及当不能与辅酶作用的突变形式表达时的影响。 这一合作努力以多种方式回应了两个区域英才中心根据“新机遇”倡议提出的申请。首先，我们将深入了解A类病原体，对于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，而且还将确定潜在的关键靶点(S)，用于未来设计 对病毒最丰富的传染性形式的干预。其次，这项提案的资金将允许主要研究人员之一(Victor Hsu)与痘病毒专家(Stuart Isaacs)合作，从而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 这是一项全新的研究，合作的互补专业知识可能会加强对牛痘病毒如何与宿主相互作用的阐明。第三，这项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 R01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.