Products of the Transforming Genes of Polyomavirus

多瘤病毒转化基因产物

• 批准号: 6989675
• 负责人: BRIAN S SCHAFFHAUSEN
• 金额: $ 33.85万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-27 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6989675
• 关键词: Polyomavirus muris 1; binding proteins; gene expression profiling; guanosinetriphosphatase activating protein; guanosinetriphosphatases; human tissue; laboratory mouse; laboratory rabbit; metastasis; neoplasm /cancer genetics; neoplastic transformation; osteopontin; phosphatidylinositol 3 kinase; protein sequence; tumor antigens; viral carcinogenesis; virus genetics

Murine polyomavirus provides a valuable model for neoplastic transformation. The virus causes a broad spectrum of tumors in animals. This means that leads developed in the laboratory can be readily tested in the host. Studies on polyoma have repeatedly provided insight into basic mechanisms of cellular growth regulation. Tyrosine kinase signaling and phosphatidylinositol 3-kinase (PI3K) signaling are two fundamental mechanisms uncovered from studies on polyoma. Middle T antigen (MT) is the major transforming protein of polyomavirus. It is always necessary, and often sufficient, for cellular transformation by polyoma. This subproject will extend our knowledge of MT and its signaling based on leads developed in the previous project period. There are 4 specific aims. 1) We have shown that MT activates the small GTPase Ral. We will examine how MT activates Ral. We will test the role of Ral in MT transformation. Since Ral functions in trafficking processes including secretion, we will test MT function there. 2) MT induces osteopontin, a secreted protein thought to be linked to transformation and metastasis. We will determine how MT regulates osteopontin. We will determine the role of osteopontin in MT transformation by examining tumor profiles. Using breast cancer and hemangioma models, we will test the role of osteopontin in metastasis and in cell recruitment. We will initiate structural analysis of MT using NMR to determine the structure of the portion of MT that interacts with downstream targets such as Shc and PI3K. We will then map the interaction surface between MT and p85 of PI3K or Shc. Finally, there is strong genetic evidence suggesting that there are undiscovered MT binding proteins. We will use tandem affinity purification and mass spectrometry to identify them. We will determine the MT sequences required for their interaction. We will test their function in MT assays for transformation.

鼠多瘤病毒为肿瘤转化提供了宝贵的模型。该病毒在动物中引起广泛的肿瘤。这意味着在实验室中开发的铅可以在宿主中很容易测试。多瘤的研究反复洞悉了细胞生长调节的基本机制。酪氨酸激酶信号传导和磷脂酰肌醇3-激酶（PI3K）信号传导是从多瘤研究中发现的两种基本机制。中T抗原（MT）是多瘤病毒的主要转化蛋白。始终是必要的，而且通常足够，用于通过多瘤的细胞转化。该副本将根据上一个项目期间开发的潜在客户扩展我们对MT及其信号的知识。有4个具体目标。 1）我们已经表明MT激活了小的GTPase RAL。我们将研究MT如何激活RAL。我们将测试RAL在MT转化中的作用。由于RAL在包括分泌在内的运输过程中的功能，因此我们将在那里测试MT功能。 2）MT诱导骨桥蛋白，这是一种分泌的蛋白质，被认为与转化和转移有关。我们将确定MT如何调节骨桥蛋白。我们将确定骨桥的作用 在MT转化中，通过检查肿瘤谱。使用乳腺癌和血管瘤模型，我们将测试骨桥蛋白在转移和细胞募集中的作用。我们将使用NMR对MT进行结构分析，以确定与SHC和PI3K等下游目标相互作用的MT部分的结构。然后，我们将绘制PI3K或SHC的MT和P85之间的相互作用表面。最后，有强烈的遗传证据表明存在未发现的MT结合蛋白。我们将使用串联亲和力纯化和质谱法来识别它们。我们将确定其相互作用所需的MT序列。我们将测试它们在MT分析中的功能以进行转换。