ID OF NOVEL CELLULAR & VIRAL PARTNERS OF THE ADENOVIRUS L4 100K PROTEIN

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们实验室的工作涉及使用腺病毒作为潜在的溶瘤剂，以及研究哺乳动物细胞翻译调控的工具。腺病毒溶瘤疗法的原型是一种突变的腺病毒-ONYX-015-它被设计成在肿瘤细胞中复制，而不是在正常细胞中复制。然而，在某些肿瘤细胞中，0 NYX-015不能复制，这是因为它不能表达L4 100 K蛋白，L4 100 K蛋白参与抑制宿主蛋白合成和晚期病毒信息的特异性翻译，这是一个对生产性腺病毒感染至关重要的过程。因此，我研究L4- 100 k的调节机制，以及这种蛋白质如何介导宿主蛋白质合成关闭。它是一种多功能蛋白质，具有RNA结合活性，已知可与翻译起始复合物结合并阻止细胞信息的帽依赖性翻译，从而促进可以以帽非依赖性方式发生的病毒特异性翻译。该项目的这一部分涉及TAP纯化和质谱，旨在鉴定100 k的新细胞伴侣，这是理解这种蛋白质功能的关键，应该让我们更多地了解如何设计更有效的溶瘤载体，并可能提供对哺乳动物细胞中翻译控制机制的见解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Work in our lab involves using adenovirus as a potential oncolytic agent, as well as a tool to study regulation of translation in mammalian cells. The prototype of adenoviral oncolytic therapy is a mutant adenovirus -ONYX-015- which was designed to replicate in tumor cells but not in normal cells. However, in certain tumor cells, 0NYX-015 fails to replicate and this was shown to be because it fails to express the L4 100K protein, which is involved in inhibition of host protein synthesis and specific translation of late viral messages, a process which is critical for a productive adenovirus infection. Therefore, I study the mechanism by which L4-100k is regulated, and how this protein mediates host protein synthesis shutoff. It is a multifunctional protein, has RNA binding activity and is known to bind to the Translation initiation complex and prevent cap-dependent translation of cellular messages, thus promoting virus-specific translation which can occur in a cap-independent manner. This part of the project, which involves TAP purification and Mass spec, aims at identifying novel cellular partners of 100k, which is key to understanding the function of this protein and should give us more insights into how to design more effective oncolytic vectors, and may provide insights into the mechanism of translational control in mammalian cells.