Molecular dissection studies of mammalian reovirus proteins using reverse genetics.

哺乳动物呼肠孤病毒在我们理解重要的生物学机制方面发挥了重要作用：病毒入侵、核酸和蛋白质的合成、病毒进化等。除了它作为一种基础研究工具的兴趣之外，现在还设想了这种迷人的病毒的可能应用。事实上，一些病毒，如呼肠孤病毒，可能会成为我们的盟友，而不仅仅是敌人。它能够感染人类以及大量动物物种，而不会对人类造成疾病，这进一步促使人们从这种病毒中开发出“病毒治疗剂”。例如，现在已经确定呼肠孤病毒优先感染和杀死癌细胞。可以进一步设想将该病毒开发为基因转移载体。相反，在过去几年中报告了新的呼肠孤病毒致病株，这表明新出现的呼肠孤病毒可能对人类和动物构成新的威胁。在过去的25年里，实验室成员对我们对病毒的了解做出了重大贡献。分子生物学、生物化学和经典遗传学方法对剖析许多病毒蛋白质的结构和功能特别有用。2007年，我们见证了一项重大进展，一种新的、强大的基于质粒的反向遗传学方法的发展，这种方法可以利用基因工程来产生各种病毒突变体。这是一项重大进展，现在可以在病毒复制周期的背景下研究病毒蛋白质的各种变化的影响。我们是少数几个愿意采用这种有希望的方法的实验室之一。在过去的几年里，它被广泛用于生产和研究具有潜在有趣特性的新的呼肠孤病毒变体。在本文提出的研究中，将研究三种不同的病毒衣壳蛋白：它们发挥不同的作用，但尚不完全清楚，特别是在病毒进入、病毒核酸合成和控制宿主细胞对感染的反应方面。除了在基础知识方面的净收益，我们的研究计划还受到旨在开发和优化病毒应用以及实现对可能出现的病原体的更好控制的长期目标的推动。

Mammalian reovirus has played an important role in our understanding of important biological mechanisms: virus entry, synthesis of nucleic acids and proteins, virus evolution, and more. In addition to its interest as a fundamental research tool, possible applications are now envisaged for this fascinating virus. In fact, some viruses, such as reovirus, could become our allies rather than just enemies to fight. Its ability to infect human beings, as well as a large number of animal species, without causing disease in humans, is a further incentive to develop “virotherapeutic” agents from this virus. For example, it is now well established that reovirus preferentially infects and kills cancer cells. It can be further envisaged developing the virus as a gene-transfer vector. On the contrary, new pathogenic isolates of reovirus have been reported in the last few years, suggesting the possibility of emerging reoviruses that could represent new threats to humans and animals. During the last 25 years, members of the laboratory have significantly contributed to our understanding of the virus. Molecular biology, biochemistry, and classical genetics approaches, have been especially useful to dissect the structure and function of many viral proteins. In 2007, we witnessed a major progress with the development of a new and powerful method of plasmid-based reverse genetics which could be used to produce various viral mutants, using genetic engineering. This was a major progress, now allowing to study the effect of various changes to the viral proteins in the context of the viral replicative cycle. We were among the few laboratories to readily adopt this promising approach. It has been used extensively in the last few years to produce and study new reovirus variants that possess potentially interesting properties. In the studies proposed herein, three different viral capsid proteins will be studied: these play different, yet incompletely understood, roles, especially in virus entry, synthesis of viral nucleic acids and control of the host-cell response to infection. In addition to the net gains in fundamental knowledge, our research program is motivated by longer-term objectives aimed at developing and optimizing applications of the virus as well as achieving better control of the possible emerging pathogens.