A novel protein regulating thymocyte development

调节胸腺细胞发育的新型蛋白质

• 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
• 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

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.

首席调查员/项目主任(最后、第一、中间)：Gascoigne，Nicholas R.J.1 R01 AI073870-01A2 一个新的基因/蛋白已被发现，它主要在胸腺细胞发育过程中表达。基因敲除的小鼠表明，它是正常胸腺细胞阳性选择所必需的。初步数据表明，它与磷脂酶C-伽马(PLC-伽马)和ITK相互作用，这两种酶对早期 通过TCR以及毛细血管扩张性共济失调突变(ATM)的信号传递阶段，ATM是细胞周期检查点的主要调节因子，用于测试DNA损伤。这项应用旨在确定这种蛋白质的功能，并确定它如何与发育中的胸腺细胞的信号级联相互作用。在正选择信号之前和期间，基因敲除胸腺细胞基因表达谱的变化将是 确认身份。该蛋白质的潜在功能区将被识别和分析，以了解它们在胸腺细胞发育中的作用，包括核定位序列、ATM磷酸化的位点、SH3结合位点等。将使用FRET显微镜和荧光互补技术来研究与ITK和PLC-伽马相互作用的动力学。诱导细胞死亡或分化 发展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着这些过程，因此这个项目将揭示这种新蛋白质是如何发挥作用的。