A novel protein regulating thymocyte development

调节胸腺细胞发育的新型蛋白质

• 批准号: 7532691
• 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/7532691
• 关键词: Affect; Antigens; Autoimmunity; Binding; Binding Sites; Bone Marrow Stem Cell; Cell Cycle; Cell Cycle Checkpoint; Cell Death Induction; Cell Differentiation process; Cells; DNA Damage; DNA Repair; DNA Sequence Rearrangement; Data; Defect; Drops; Enzymes; Family; Fluorescence; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Funding; Gene Expression; Gene Proteins; Genetic Transcription; Goals; Grant; Hybrids; Immune response; Immune system; Knock-out; Knockout Mice; Microscopy; Molecular Profiling; Motion; Mouse Strains; Mus; Mutation; Nuclear; Nuclear Localization Signal; PH Domain; Phospholipase C; Phosphorylation Site; Phosphotransferases; Play; Principal Investigator; Process; Protein Region; Proteins; Research Project Grants; Role; SH3 Domains; Serine/Threonine Phosphorylation; Signal Pathway; Signal Transduction; Site; Staging; Study Section; T-Cell Activation; T-Cell Development; T-Lymphocyte; Testing; Thymocyte Development; Tyrosine; Tyrosine Phosphorylation Site; Upper arm; Yeasts; ataxia telangiectasia mutated protein; base; in vivo; member; novel; phospholipase C gamma; polyproline; programs; protein function; reconstitution; sound; 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.

首席研究员/项目总监（最后、第一、中间）：Gascoigne、Nicholas R.J. 1 R01 AI073870-01A2 已鉴定出主要在胸腺细胞发育过程中表达的新基因/蛋白质。基因敲除小鼠表明，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-gamma (PLC-gamma) 和 Itk 相互作用，这两种酶对于早期发育至关重要。 通过 TCR 以及毛细血管扩张共济失调突变 (ATM) 进行信号传导的各个阶段，ATM 是测试 DNA 损伤的细胞周期检查点的主要调节因子。该应用旨在鉴定该蛋白质的功能，并确定其如何与发育中的胸腺细胞中的信号级联反应相互作用。在正选择信号传导之前和期间，敲除胸腺细胞中基因表达谱的变化将是 确定。该蛋白质的潜在功能区域将被鉴定并分析其在胸腺细胞发育中的作用，包括核定位序列、ATM 磷酸化位点、SH3 结合位点等。将使用 FRET 显微镜和荧光互补研究与 Itk 和 PLC-gamma 相互作用的动态。诱导细胞死亡或分化 发育 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可以调节这些过程，因此该项目将揭示这种新蛋白质的功能。