Late Events in Retrovirus Assembly

描述(由申请人提供)：在这份竞争性的续签申请中，我们请求支持我们对逆转录病毒组装中的晚期事件的研究。在上一个资助周期中，我们鉴定并鉴定了几种通过病毒晚萌发(L)结构域被招募到逆转录病毒组装部位的细胞蛋白。这些蛋白质或者是(I)E类空泡蛋白分选(VPS)途径的组成部分，该途径通常参与内小体和多囊泡小体(MVB)的操纵；或者(Ii)泛素连接酶。对于这些因素中的几个，我们能够证明在传染性逆转录病毒颗粒的释放中起关键作用。然而，一些机制问题仍然存在，最近我们发现了更多的细胞因子，它们似乎非常可能参与逆转录病毒颗粒的构建和释放的后期阶段，和/或内体/MVB的生物发生。在下一个资助期间，我们将试图通过追求两个特定的目标来阐明先前定义的和新发现的细胞因子如何参与逆转录病毒组装的后期阶段和相关的细胞功能。具体目标1是了解特定的内体蛋白如何结合到HIV-1颗粒中，以及它们在产生感染性病毒粒子中的作用。具体目标2是确定在前一资金时期发现的泛素、泛素连接酶和新型泛素连接酶结合蛋白在逆转录病毒萌芽和内吞蛋白运输中的作用。这两个目标的实现应该会导致我们朝着了解细胞和病毒因素如何在构建完整的、有感染性的逆转录病毒颗粒中合作的长期目标取得重大进展。了解这些事件可能会为干预艾滋病毒和其他包膜病毒引起的疾病的治疗过程提供新的机会。公共卫生相关性：许多包膜病毒，例如HIV-1，是威胁生命的感染的原因。为了在细胞间传播，它们利用细胞蛋白促进准确组装的病毒颗粒的释放。了解这个系统是如何工作的，可以提供进行靶向治疗干预的机会。

DESCRIPTION (provided by applicant): In this competing renewal application we request support for our studies on late events in retrovirus assembly. In the previous funding cycle, we identified and characterized several cellular proteins that are recruited to sites of retrovirus assembly by viral late budding (L)-domains. These proteins are either (i) components of the class E vacuolar protein sorting (VPS) pathway that is normally involved in the manipulation of endosomes and multivesicular bodies (MVB) or (ii) ubiquitin ligases. For several of these factors we were able to demonstrate a critical role in the release of infectious retroviral particles. Several mechanistic questions remain, however, and recently we have identified additional cellular factors that seem very likely to be involved in the late stages of the construction and release of retrovirus particles, and/or endosome/MVB biogenesis. In the next grant period, we will attempt to elucidate how previously defined and newly identified cellular factors participate in the late stages of retrovirus assembly and related cellular functions by pursuing two specific aims. Specific Aim 1 is to understand how specific endosomal proteins are incorporated into HIV-1 particles and their role in generating infectious virions. Specific aim 2 is to determine the role of ubiquitin, ubiquitin ligases and novel ubiquitin ligase binding proteins, discovered during the previous funding period, in retroviral budding and trafficking of endocytosed proteins. Execution of these two aims should result in significant progress toward our long term goal of understanding how cellular and viral factors cooperate in the construction of complete, infectious retroviral particles. Understanding these events could potentially provide new opportunities to interfere in these processes for the treatment of diseases caused by HIV and other enveloped viruses. Public health relevance: Many enveloped viruses, for example HIV-1, are responsible for life threatening infections. To spread from cell to cell they make use of cellular proteins to facilitate the release of accurately assembled virus particles. Understanding how this system works could give opportunities to intervene with targeted therapeutics.