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％的细胞磷酸磷酸/苏氨酸磷酸酶活性，蛋白质磷酸酶2a （PP2A）调节几乎所有细胞信号通路。 pp2a> 80种异聚体，组成催化C亚基和与Aα结合的许多调节B亚基之一（丰富90％）或（10％ Aβ亚基支架。我们的前提是使用Aβ支架的蛋白质磷酸酶2a（PP2A）为对于控制正常和癌细胞的表型的根本重要。此外，研究多瘤病毒反复给予了对生长控制的新颖见解，将是无价的了解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/siRNAAβ敲低确认即使在没有病毒的情况下，对于癌症重要的途径也很重要。 AKT和C-SRC信号都是由Aβ调节。此外，人肺，乳腺癌和结肠癌在Aβ中显示出改变，表明 Aβ-PP2A介导的信号传导与癌症有关。在Aβ上完成的工作很少，所以有一个迫切需要研究其功能。在AIM 1中，我们将在功能调查中使用基于广泛的技术并整合这些方法以识别正常和转化细胞中Aβ改变的途径。通过RNA-SEQ，磷酸蛋白质组学分析和NMR代谢组学识别途径的表达分析由PP2AAβ特别针对。对照与表达MUPYV MT或ST的单元的对照进行比较将告知我们是否抑制Aβ活性和/或将其重定向到新靶标。目标2，专注于 Aβ将确定Aβ/ST结构。将确定PP2A B亚基和其他结合Aβ的靶标。 Aβ的遗传分析将发现负责其独特表型的序列。在AIM 3Aβ中将检查酪氨酸激酶信号传导的调节，以了解C-SRC控制的机制和TO 确定Aβ如何通过酪氨酸磷酸酶进行广泛控制酪氨酸磷酸化。英石将确定缺陷Aβ结合的突变体，以测试Aβ在控制细胞表型中的作用。最后，我们将使用条件敲除技术确认Aβ在HER2/NEU和MT肿瘤发生中的作用。呢