A novel protein regulating thymocyte development

调节胸腺细胞发育的新型蛋白质

基本信息

批准号：
7842644
负责人：
NICHOLAS R GASCOIGNE
金额：
$ 47.48万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-15 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7842644
关键词：
Affect Autoimmunity Binding Binding Sites Biology Bone Marrow Stem Cell Cell Cycle Cell Cycle Checkpoint Cell Death Induction Cells DNA Damage DNA Repair DNA Sequence Rearrangement Data Defect Enzymes Family Fluorescence Fluorescence Microscopy Fluorescence Resonance Energy Transfer Gene Expression Gene Proteins Genes Genetic Transcription Goals Hybrids Immune response Immune system Interferons Knock-out Knockout Mice Mature T-Lymphocyte Mature Thymocyte Mediating Memory Methods Microscopy Molecular Profiling Motion Mus Mutation Nuclear Nuclear Localization Signal Organ PH Domain Phospholipase Phosphorylation Site Phosphotransferases Principal Investigator Process Protein Region Proteins Role SH3 Domains Serine/Threonine Phosphorylation Signal Pathway Signal Transduction Site Staging T-Cell Activation T-Cell Development T-Lymphocyte Tamoxifen Testing Thymocyte Development Time Tyrosine Tyrosine Phosphorylation Site Yeasts ataxia telangiectasia mutated protein base in vivo member novel phospholipase C gamma polyproline programs promoter protein function reconstitution tandem mass spectrometry thymocyte

项目摘要

Principal Investigator/Program Director (Last, first, middle): Gascoigne, Nicholas R.J. 1 R01 AI073870-01A2 A novel gene/protein has been identified which is expressed predominantly during thymocyte development. A knockout mouse shows that it is required for normal thymocyte positive selection. Preliminary data indicate that it interacts with phospholipase C-gamma (PLC-gamma) and Itk, both enzymes crucial to early stages in signaling via the TCR, as well as with Ataxia telangiectasia mutated (ATM), a master regulator of cell cycle checkpoints that tests for DNA damage. This application aims to identify the function of this protein and to determine how it interacts with the signaling cascades in developing thymocytes. Changes in gene expression profiles in the knockout thymocytes before and during positive selection signaling will be identified. Potential functional regions of the protein will be identified and analyzed for their role in developing thymocytes, including a nuclear localization sequence, a site for phosphorylation by ATM, an SH3-binding site and others. The dynamics of the interaction with Itk and PLC-gamma will be investigated using FRET microscopy and fluorescence complementation. The induction of cell death or differentiation in developing T cells is crucial for avoiding autoimmunity, and for building a functional immune system. This newly discovered protein appears to regulate these processes, thus this project will uncover how this novel protein functions.

首席研究员/计划主任（最后，第一，中间）：加斯科尼，尼古拉斯·R·J。 1 R01 AI073870-01A2 已经鉴定出一种新型的基因/蛋白质，该基因/蛋白在胸腺细胞发育过程中主要表达。敲除小鼠表明正常胸腺细胞阳性选择是必需的。初步数据表明，它与磷脂酶C-gamma（PLC-Gamma）和ITK相互作用，这都是至关重要的酶通过TCR以及temaxia telangiectia突变（ATM）的信号传导的阶段，这是测试DNA损伤的细胞周期检查点的主调节器。该应用旨在识别该蛋白质的功能，并确定其与发育中的胸腺细胞中信号级联反应的相互作用。在阳性选择信号之前和期间，基因表达谱的变化将是确定。将鉴定并分析蛋白质的潜在功能区域，以发挥其在开发胸腺细胞中的作用，包括核定位序列，ATM，SH3结合位点等的ATM磷酸化位点。将使用FRET显微镜和荧光互补研究与ITK和PLC-GAMMA相互作用的动力学。诱导细胞死亡或分化开发T细胞对于避免自身免疫和建立功能性免疫系统至关重要。这种新发现的蛋白质似乎调节了这些过程，因此该项目将发现这种新型蛋白质的功能。