Viral modulation of cellular RNA alternative splicing

我们研究计划的长期目标是了解病毒蛋白的分子和功能作用。我们的实验室对参与基因表达和RNA代谢调节的病毒蛋白特别感兴趣，因为我们对病毒蛋白如何影响这些过程的理解仍然存在重大差距。了解病毒如何与宿主细胞相互作用，以及病毒最终如何修饰细胞基因的表达，对于充分了解病毒复制至关重要。目前的研究重点是病毒蛋白对选择性剪接的影响，这是导致蛋白质多样性增加的关键RNA加工步骤。真核生物的早熟rna在输出到细胞质之前经过广泛的修饰，在那里它们被翻译成蛋白质。在这些成熟过程中，RNA剪接是至关重要的，它允许从早熟RNA转录物中去除内含子。然而，一些外显子，部分外显子和/或内含子也可以通过称为选择性剪接（AS）的过程保留或排除在成熟转录本中。在功能上，AS通过对成熟mrna中所包含的外显子的差异选择来改变所产生蛋白质的组成，从而影响细胞蛋白质组。在过去的几年中，我们的团队和其他人已经证明，病毒感染会导致受感染细胞的AS景观发生剧烈变化。这些变化具有重塑感染细胞蛋白质组的重要潜力，从而调节病毒与宿主细胞之间的相互作用。为了更好地了解病毒与宿主的相互作用，需要更好地了解病毒如何调节细胞AS。我们的研究假设是特定的病毒蛋白参与了细胞AS的调节。为了验证这一假设，我们提出了三个具体目标：1-鉴定参与细胞AS调节的病毒蛋白。2-剖析细胞AS调节的分子机制。3-修改病毒感染细胞的剪接程序，以确定AS在病毒感染中的作用。为了实现这些目标，我们将使用不同的病毒作为模型（呼肠孤病毒、西尼罗河病毒和Epstein-Barr病毒）。结合分子和转录组学工具，我们将破译病毒调节细胞AS的机制。研究生和本科生都将是我们研究项目的驱动力。通过他们在分子生物学和病毒学方面的训练，他们将继续为该领域做出原创性贡献。显然，我们的首要任务是利用国家科学研究委员会的资助，为有兴趣从事有回报的研究事业的学生提供优秀的培训。

The long-term objective of our research program is to understand the molecular and functional roles of viral proteins. Our laboratory is especially interested in viral proteins that are involved in the regulation of gene expression and RNA metabolism because there are still significant gaps in our understanding of how viral proteins affect these processes. Understanding how viruses interact with their host cells, and ultimately how viruses modify the expression of cellular genes, is essential to fully understand viral replication. The current research program focuses on the influence of viral proteins on alternative splicing, a pivotal RNA processing step which results in increased protein diversity. Eukaryotic pre-mature RNAs undergo extensive modifications before being exported to the cytoplasm where they are translated into proteins. Amongst those maturation processes, RNA splicing is vital by allowing the removal of introns from pre-mature RNA transcripts. However, some exons, portions of exons and/or introns can also be retained or excluded from the mature transcript by a process called alternative splicing (AS). Functionally, AS impacts the cellular proteome by changing the composition of resulting proteins through differential choice of exons to be included in mature mRNAs. During the past few years, our group and others have demonstrated that viral infection leads to drastic changes in the AS landscape of infected cells. These changes have an important potential to reshape the proteome of infected cells thereby modulating the interplay between viruses and their host cells. A better understanding of how viruses modulate cellular AS is needed to gain a better global understanding of virus-host interactions. Our research hypothesis is that specific viral proteins are involved in the modulation of cellular AS. To verify this hypothesis, we propose three specific aims: 1- To identify viral proteins involved in the modulation of cellular AS. 2- To dissect the molecular mechanisms responsible for the modulation of cellular AS. 3- To modify the splicing program of virus-infected cells to identify the roles of AS during viral infection. To achieve these goals, we will use different viruses as models (reovirus, West Nile virus, and Epstein-Barr virus). Using a combination of molecular and transcriptomic tools, we will decipher the mechanisms used by viruses to modulate cellular AS. Both graduate and undergraduate students will be the driving force in our research program. Through their training in molecular biology and virology, they will continue to make original contributions to the field. Clearly, it is a priority for us to use the NSERC grant to provide excellent training for students interested in rewarding careers in research.