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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们实验室的工作包括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变异型腺病毒--Onyx-015，它被设计成在肿瘤细胞中复制，但在正常细胞中不复制。然而，在某些肿瘤细胞中，0NYX-015不能复制，这被证明是因为它不能表达L4 100K蛋白，该蛋白参与抑制宿主蛋白质合成和晚期病毒信息的特定翻译，这一过程对腺病毒的有效感染至关重要。因此，我研究了L4-100k的调控机制，以及该蛋白是如何介导宿主蛋白合成停止的。它是一种多功能蛋白质，具有RNA结合活性，已知能与翻译起始复合体结合，阻止细胞信息的帽依赖翻译，从而促进病毒特异性翻译，这种翻译可以以帽不依赖的方式发生。该项目的这一部分包括TAP纯化和质谱仪，旨在寻找100K的新细胞伙伴，这是理解该蛋白功能的关键，应该会给我们提供更多关于如何设计更有效的溶瘤载体的见解，并可能为哺乳动物细胞中翻译调控的机制提供见解。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.