SIGNAL TRANSDUCTION PATHWAYS AND POLYOMA MIDDLE T

信号转导途径和多发性中 T

• 批准号: 6269423
• 负责人: BRIAN S SCHAFFHAUSEN
• 金额: $ 27.41万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-06 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6269423
• 关键词: G protein; Polyomavirus; alcohol phosphotransferase; biological signal transduction; cell growth regulation; chemical binding; enzyme activity; host organism interaction; immunoprecipitation; laboratory mouse; laboratory rabbit; laboratory rat; mutant; neoplastic transformation; oncoproteins; phosphorylation; protein sequence; protein structure function; site directed mutagenesis; tissue /cell culture; transforming virus; tumor antigens; viral carcinogenesis; virus antigen; virus protein

The overall goals are to understand pathways of cell growth regulation and to understand how polyomavirus subverts them to produce transformation. The work proposed here concerns signaling pathways employed for transformation by polyoma middle T antigen. It emphasizes the enzyme phosphatidylinositol-3'-kinase. Understanding PI-3'-kinase is essential not only because it appears critical for middle T transformation, but also because this enzyme has been implicated in growth regulation by other transforming genes and growth factor receptors. In this enzyme, the P110 subunit has catalytic activity, while the p85 subunit provides regulation. The p85 subunit contains SH2, SH3 and bcr domains. These elements are found in a wide variety of proteins involved in signal response. The results obtained for PI-3'- kinase will therefore be of general importance. Our specific goals are: A. Studies on regulation of PI-3'-kinase and its role in growth and transformation 1. Studies on sites of serine phosphorylation 2. Analysis of the effect of blocking PI-3'-kinase activation 3. Establishment of connections between PI-3'-kinase and other growth regulatory pathways B. Studies related to SH2 function 1. Random mutagenesis of SH2s to determine structure/function relationships 2. Determination of whether middle T binds simultaneously to more than one SH2 3. Test of the MXM hypothesis of SH2 specificity to see if a designed middle T mutant can bind a p85 with an appropriately altered specificity. 4. Determination of whether an additional SH2 interaction might be important for determining tumor profiles of hamster middle T. C. Studies related to SH3 function 1. Mutagenesis of the SH3 of PI-3'-kinase 2. Identification of proteins that bind p85 SH3 3. Mutagenesis of middle T in the region of dl 1015 4. Screening for interactions between middle T and other SH3s D. Studies of the interactions among small G-proteins, PI-3'-kinase and middle T 1. Determination of PI-3'-kinase rac/rho/cdc 42 interactions 2. Analysis of the role of rac/rho in polyoma transformation using dominant negatives.

总体目标是了解细胞生长调节的途径 并了解多瘤病毒如何颠覆它们以生产 转型。 这里提出的工作涉及信号通路 用于通过多瘤中间T抗原转化的用于转化。 它强调 酶磷脂酰肌醇-3'-激酶。 了解Pi-3'-激酶 这是必不可少的，不仅是因为它似乎对中间 转型，也是因为这种酶已与 其他转化基因和生长因子的生长调节 受体。 在这种酶中，p110亚基具有催化活性， p85亚基提供了调节。 p85亚基包含SH2， SH3和BCR域。 这些元素在各种各样的 涉及信号反应的蛋白质。 PI-3'-获得的结果 因此，激酶将非常重要。 我们的具体目标是： A.对PI-3'-激酶调节及其在生长中的作用的研究和 转型 1。丝氨酸磷酸化部位的研究 2。分析阻断PI-3'-激酶激活的效果 3。建立Pi-3'-激酶与其他增长之间的联系 监管途径 B.与SH2功能有关的研究 1。SH2S的随机诱变以确定结构/功能 关系 2。确定中间t是否同时绑定比 一个SH2 3。SH2特异性的MXM假设的测试以查看是否设计 中间突变体可以通过适当改变的p85结合 特异性。 4。确定是否可能是其他SH2相互作用 对于确定仓鼠中间T的肿瘤特征很重要。 C.与SH3功能有关的研究 1。pi-3'-激酶SH3的诱变 2。鉴定结合p85 SH3的蛋白质 3。dl 1015区域中间T的诱变 4。筛选中间T和其他SH3之间的相互作用 D.研究小G蛋白之间的相互作用，PI-3'-激酶和 中间 1。确定PI-3'-激酶RAC/RHO/CDC 42相互作用 2。分析RAC/RHO在使用Polyoma转化中的作用 主要的负面因素。