PROJECT 3: Pathways Controlled by PP2A A-Beta in Normal, Transformed and Tumor Cells

项目 3：正常细胞、转化细胞和肿瘤细胞中 PP2A A-Beta 控制的通路

基本信息

批准号：
10227784
负责人：
BRIAN S SCHAFFHAUSEN
金额：
$ 35.42万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10227784
关键词：
1-Phosphatidylinositol 3-Kinase Affect Amyloid beta-Protein Antigens Attention Binding Breast Cell Survival Cells Colon Carcinoma Complex Critical Pathways Data ERBB2 gene Gene Expression Gene Proteins Genetic Structures Growth Growth Factor Receptors Health Human Knock-out Learning Malignant Neoplasms Malignant neoplasm of lung Mediating Merkel Cells Metabolic Metabolism Modeling Mus Mutagenesis Mutation Normal Cell Oncogenic Oncogenic Viruses Oncoproteins PTPN1 gene Pathway interactions Phenotype Phosphoric Monoester Hydrolases Phosphoserine Polyomavirus Protein Dephosphorylation Protein Isoforms Protein Tyrosine Kinase Protein Tyrosine Phosphatase Protein phosphatase Proteomics RNA analysis Reagent Regulation Role SRC gene Signal Pathway Signal Transduction Small Interfering RNA Structure Surveys Swiss 3T3 Cells Technology Testing Threonine Tissues Transgenic Mice Tyrosine Phosphorylation Viral Virus Work X ray diffraction analysis base cancer cell cell transformation cell type cellular targeting conditional knockout embryonic stem cell established cell line experimental study genetic analysis glucose metabolism insight knock-down malignant breast neoplasm metabolomics mouse polyomavirus mutant neoplastic cell novel novel strategies phosphoproteomics prevent protein function scaffold src-Family Kinases transcriptome sequencing tumor tumorigenesis

项目摘要

Responsible for up to 50% of cellular phosphoserine/threonine phosphatase activity, protein phosphatase 2A (PP2A) regulates almost all cell signaling pathways. PP2A comes as > 80 kinds of heterotrimers, consisting of a catalytic C subunit and one of many regulatory B subunits bound to an Aα (90% abundant) or (10% abundant) Aβ subunit scaffold. Our premise is that protein phosphatase 2A (PP2A) using the Aβ scaffold is fundamentally important for controlling phenotypes of both normal and cancer cells. Moreover, study of polyomaviruses, which have repeatedly given novel insights into growth control, will be invaluable to understand Aβ function. Our studies of murine polyomavirus (MuPyV) ST/MT already demonstrate the importance of Aβ to survival, differentiation and transformation. Our sh/siRNA Aβ knockdowns of confirm its importance, even in the absence of virus, to pathways important in cancer. Both Akt and c-Src signaling are regulated by Aβ. In addition, human lung, breast and colon cancers show alterations in Aβ, suggesting that Aβ-PP2A-mediated signaling is relevant to cancer. Very little work has been done on Aβ, so there is a pressing need to study its function. In Aim 1 we will use broad-based technologies in a survey of functions and integrate these approaches to identify pathways altered by Aβ in normal and transformed cells. Expression analysis by RNA-seq, phosphoproteomic analysis, and NMR metabolomics will identify pathways specifically targeted by PP2A Aβ. Comparisons of controls with cells expressing MuPyV MT or ST will inform us whether the oncoproteins are inhibiting Aβ activity and/or redirecting it to new targets. Aim 2, focusing on Aβ, will determine the Aβ/ST structure. PP2A B subunits and other targets that bind Aβ will be determined. Genetic analysis of Aβ will uncover sequences responsible for its unique phenotype(s). In Aim 3 Aβ regulation of tyrosine kinase signaling will be examined to learn the mechanism of c-Src control and to determine how Aβ broadly controls tyrosine phosphorylation, perhaps via tyrosine phosphatases. ST mutants defective for Aβ binding will be identified to test the role of Aβ in controlling cell phenotype. Finally, we will confirm the role of Aβ in Her2/neu and MT tumorigenesis using conditional knockout technology. !

负责高达 50% 的细胞磷酸丝氨酸/苏氨酸磷酸酶活性（蛋白磷酸酶 2A） (PP2A) 调节几乎所有的细胞信号通路，由超过 80 种异源三聚体组成。催化 C 亚基和与 Aα 结合的许多调节 B 亚基之一（丰度 90%）或（10%）丰富的）Aβ亚基支架我们的前提是使用Aβ支架的蛋白磷酸酶2A（PP2A）是对于控制正常细胞和癌细胞的表型至关重要。多瘤病毒多次为生长控制提供了新的见解，对于了解 Aβ 功能。我们对鼠多瘤病毒 (MuPyV) ST/MT 的研究已经证明了 Aβ 对生存、分化和转化的重要性我们的 sh/siRNA Aβ 敲低证实了它的重要性。即使在没有病毒的情况下，Akt 和 c-Src 信号传导对于癌症中的重要通路也很重要。此外，人类肺癌、乳腺癌和结肠癌中 Aβ 也存在变化，这表明 Aβ-PP2A 介导的信号传导与癌症相关，目前对 Aβ 的研究还很少，因此存在一个问题。在目标 1 中，我们将使用广泛的技术来研究其功能。并整合这些方法来识别正常细胞和转化细胞中 Aβ 改变的途径。通过 RNA-seq、磷酸化蛋白质组学分析和 NMR 代谢组学进行表达分析将确定通路 PP2A Aβ 特异性靶向对照与表达 MuPyV MT 或 ST 的细胞的比较将提供信息。我们研究癌蛋白是否抑制 Aβ 活性和/或将其重定向至新目标，重点是目标 2。 Aβ 将确定 Aβ/ST 结构，并确定结合 Aβ 的其他靶标。 Aβ 的遗传分析将揭示导致其独特表型的序列 In Aim 3 Aβ。将检查酪氨酸激酶信号传导的调节，以了解 c-Src 控制机制并确定 Aβ 如何广泛控制酪氨酸磷酸化，可能是通过酪氨酸磷酸酶 ST。将鉴定 Aβ 结合缺陷的突变体，以测试 Aβ 在控制细胞表型中的作用。我们将利用条件敲除技术确认Aβ在Her2/neu和MT肿瘤发生中的作用。！