Mechanisms and regulation of post-translational modifications of Id proteins

Id蛋白翻译后修饰的机制和调控

基本信息

批准号：
7738516
负责人：
Antonio Iavarone
金额：
$ 33.41万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-03 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7738516
关键词：
Address Alanine Amino Acids Arts Biochemical Genetics Biological Assay Boxing Cell Cycle Progression Cell Differentiation process Cell physiology Cells Co-Immunoprecipitations Colon Carcinoma Complex Cyclic AMP-Dependent Protein Kinases Cyclin-Dependent Kinases Development Differentiation Inhibitor Enzymes Event Family member Genes Goals Human Id2 protein In Vitro Individual Inhibitor of Differentiation Proteins Intervention Knock-in Mouse Laboratories Malignant Neoplasms Mass Spectrum Analysis Mediating Messenger RNA Missense Mutation Molecular Mus Mutation Mutation Analysis Normal Cell Oncogene Proteins Pathway interactions Phosphorylation Phosphorylation Site Phosphotransferases Play Post-Translational Protein Processing Post-Translational Regulation Process Proteins Regulation Reporting Research Personnel Role Serine Signal Pathway Signal Transduction Site Stem cells System Testing Threonine Ubiquitin Ubiquitination Whole Organism Work anaphase-promoting complex angiogenesis base cancer cell cancer therapy follow-up in vivo metaplastic cell transformation migration mouse model mutant neoplastic cell novel novel therapeutics outcome forecast overexpression protein degradation protein expression protein function retinoblastoma tumor suppressor tissue culture tumor tumor progression tumorigenesis ubiquitin ligase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Bolstered by the overexpression of Id proteins in a wide range of human tumors and the frequent correlation with a poor prognosis, several investigators have suggested that Id proteins play key roles in the initiation and progression of cancer. Our own work identified the Retinoblastoma tumor suppressor pathway as a network that cross talks with Id proteins during development but is subverted by Id in Myc-driven cancer. However, the post-translational machinery that regulates Id proteins in normal cells is unknown. Similarly, it remains unclear whether genetic alterations leading to deregulated Id activity exist in cancer cells. Recently, we discovered that colon cancer cells acquire a missense mutation in the Id2 gene that converts Threonine-27 into the unphosphorylatable amino acid Alanine. We also found that Threonine-27 is phosphorylated by the Dyrk1 kinases in vitro and in vivo. Our Specific Aims will focus on the regulation and function of this phosphorylation event and the destruction signals that we recently identified in Id proteins. Thus, we will define the signaling pathways converging on Threonine-27 phosphorylation and elucidate how they regulate Id2 protein function. Through biochemical and genetic approaches we will also determine why tumor cells select for the Threonine-27 to Alanine Id2 mutant protein. To address this problem in the context of the whole organism, we will generate knock-in mice that conditionally express the tumor-specific mutation of Id2. Finally, we will take advantage of state-of-the-art mass spectrometry approaches developed in our laboratory to identify and characterize novel enzymes responsible for post-translational modifications of Id proteins. Through the proposed work, we aim to provide a better understanding of the post-translational regulatory machinery for Id proteins and unravel new layers of intervention targeting the alteration of this machinery in human cancer.

描述（由申请人提供）：由于 Id 蛋白在多种人类肿瘤中过度表达以及与不良预后经常相关，一些研究人员表明 Id 蛋白在癌症的发生和进展中发挥着关键作用。我们自己的工作确定了视网膜母细胞瘤肿瘤抑制通路是一个在发育过程中与 Id 蛋白相互作用的网络，但在 Myc 驱动的癌症中被 Id 颠覆。然而，正常细胞中调节 Id 蛋白的翻译后机制尚不清楚。同样，目前尚不清楚癌细胞中是否存在导致 Id 活性失调的基因改变。最近，我们发现结肠癌细胞的 Id2 基因发生错义突变，将苏氨酸 27 转化为不可磷酸化的氨基酸丙氨酸。我们还发现 Threonine-27 在体外和体内均被 Dyrk1 激酶磷酸化。我们的具体目标将集中于这种磷酸化事件的调节和功能以及我们最近在 Id 蛋白中发现的破坏信号。因此，我们将定义汇聚于 Threonine-27 磷酸化的信号通路，并阐明它们如何调节 Id2 蛋白功能。通过生化和遗传学方法，我们还将确定肿瘤细胞选择苏氨酸-27 至丙氨酸 Id2 突变蛋白的原因。为了在整个生物体的背景下解决这个问题，我们将产生有条件表达 Id2 肿瘤特异性突变的敲入小鼠。最后，我们将利用我们实验室开发的最先进的质谱方法来识别和表征负责 Id 蛋白翻译后修饰的新型酶。通过拟议的工作，我们的目标是更好地了解 Id 蛋白的翻译后调节机制，并揭示针对人类癌症中这种机制的改变的新干预措施。