MOLECULAR AND FUNCTIONAL ANALYSIS OF CD3ZETA AND ELTA IS

CD3ZETA 和 ELTA 的分子和功能分析

• 批准号: 3146264
• 负责人: LINDA KATHLEEN CLAYTON
• 金额: $ 14.4万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-01-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3146264
• 关键词: CD3 molecule; T cell receptor; T lymphocyte; apoptosis; biological signal transduction; cell death; cell differentiation; cell population study; chemical binding; gene expression; genetic manipulation; genetic promoter element; hybridomas; immunocytochemistry; laboratory mouse; laboratory rabbit; nucleic acid hybridization; polymerase chain reaction; site directed mutagenesis; thymus; tissue /cell culture; transfection

The T cell receptor (TCR) is a multisubunit complex which functions as an antigen recognition cell surface structure and a signal transduction unit on T lineage cells. The TCR consists of the heterodimeric alpha and beta subunits associated with the monomorphic gamma, delta, epsilon, zeta, and eta subunits collectively known as CD3. All subunits with the exception of eta have been cloned at cDNA and genomic levels. Until now the primary structure of eta has not been defined but recent evidence implicates an important function for this protein in TCR signalling resulting in phosphoinositide turnover as well as in the cell cycle arrest and death of T-T hybridomas upon TCR crosslinking. We have recently deduced the primary structure of the murine CD3eta from protein microsequencing and cDNA cloning. The overall goal of this project is to understand the functional role of CD3eta in T cell receptor signalling and programmed cell death, both of thymocytes and T-T hybridomas. The specific aims of these proposed studies include: 1) the characterization of CD3eta genomic organization; 2) the evaluation of CD3eta expression during thymic development using mRNA analysis of thymocytes at different stages of development and tissue section staining with specific anti-CD3eta antibodies raised against peptides synthesized from deduced CD3eta protein sequence; 3) the reconstitution of murine CD3zeta and eta in mutant cell lines in order to elucidate their respective roles in signal transduction; and 4) site directed mutagenesis of CD3zeta and eta in order to understand the structure- function basis of the differences between CD3zeta and eta in signal transduction. These studies will elucidate the role of CD3eta protein in signal transduction and may lead to an understanding of the process of cell death which results in the selection of thymocytes during development.

T细胞受体（TCR）是一种多亚基复合物， 抗原识别细胞表面结构和信号转导单元 在T细胞上。TCR由异二聚体α和β组成， 与单态的gamma、delta、zeta和 η亚基统称为CD 3。所有子单位， eta的cDNA和基因组水平克隆。到目前为止， eta的结构尚未确定，但最近的证据表明， 该蛋白在TCR信号传导中的重要功能导致 磷酸肌醇周转以及在细胞周期停滞和死亡 TCR交联后的T-T杂交瘤。我们最近推断出 来自蛋白质微测序和cDNA的鼠CD 3 eta的结构 克隆。本项目的总体目标是了解 CD 3 eta在T细胞受体信号传导和程序性细胞死亡中的作用， 胸腺细胞和T-T杂交瘤。这些建议的具体目标 研究内容包括：1）CD 3eta基因组结构的表征; 2)用mRNA评估胸腺发育过程中CD 3 eta的表达 不同发育阶段和组织的胸腺细胞分析 用特异性抗CD 3eta抗体对切片进行染色， 从推导的CD 3eta蛋白质序列合成的肽; 3） 在突变细胞系中重建鼠CD 3 ζ和eta， 阐明它们各自在信号转导中的作用;和4）位点 定向突变CD 3 ζ和eta，以了解 CD 3 zeta与eta差异的结构-功能基础 信号转导这些研究将阐明CD 3eta的作用 蛋白质在信号转导中的作用，并可能导致对 细胞死亡的过程，导致胸腺细胞的选择， 发展