The Role of Kuzbanian and TACE in T Cell Development

Kuzbanian 和 TACE 在 T 细胞发育中的作用

基本信息

批准号：
6445698
负责人：
JENNIFER O MANILAY
金额：
$ 4.42万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-02-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6445698
关键词：
T lymphocyte biological signal transduction cell differentiation cytotoxic T lymphocyte flow cytometry genetically modified animals helper T lymphocyte laboratory mouse membrane proteins metalloendopeptidases postdoctoral investigator proteolysis western blottings

项目摘要

DESCRIPTION (provided by applicant): The Notch pathway of signal transduction is important for cell fate decisions. Studies from our laboratory first suggested a role for Notch signaling in thymic development, resulting in a model in which immature thymocytes receiving a Notch signal develop into CD8+ T cells, whereas T cells not receiving a Notch signal mature into CD4+ T cells. Proteolytic processing of the Notch-1 receptor is required for generation of Notch-1-mediated signals, and Delta, a Notch ligand, undergoes proteolytic processing in Drosophila. However, whether processing of mouse Notch homologues and Notch ligands occurs and all the proteases responsible for such processing are not known. The metalloproteases Kuzbanian (Kuz) and TACE have been implicated in processing of Notch receptors and ligands. As processing of Notch is required for Notch signaling, a block in processing of Notch ligands or receptors might result in a decrease in Notch signaling. We will identify the potential Notch receptor/ligand substrates of Kuz and TACE expressed in the thymus and analyze the functional consequences of Notch receptor/ligand cleavage on thymic development. We predict that in thymocytes overexpressing a dominant-negative Kuz or TACE gene, more CD4+ T cells than CD8 T cells might develop. Reciprocally, in thymocytes overexpressing wild-type Kuz or TACE, more CD8+ T cells than CD4+ T cells will develop. These predictions will be tested in vitro and in transgenic mice. Expression and processing of the known Notch receptors and ligands in the thymus will be assessed by quantitative Western blotting, and T cell development assessed by flow cytometry. Correlations between processing and T cell development in these systems will be investigated.

描述（由申请人提供）：信号转导的Notch通路对于细胞命运的决定很重要。首先从我们的实验室进行研究提出了 Notch 信号在胸腺发育中的作用，从而导致接受 Notch 信号的未成熟胸腺细胞发育成 CD8+ T 的模型细胞，而未接收 Notch 信号的 T 细胞则成熟为 CD4+ T 细胞。 Notch-1 受体的蛋白水解过程是生成 Notch-1 介导的信号和 Notch 配体 Delta 进行蛋白水解在果蝇中加工。然而，是否处理小鼠Notch同源物和Notch配体的发生以及负责此类处理的所有蛋白酶不知道。金属蛋白酶 Kuzbanian (Kuz) 和 TACE 已被参与Notch受体和配体的加工。作为Notch的加工 Notch 信号传导、Notch 配体加工过程中的阻断或受体可能会导致 Notch 信号传导减弱。我们将确定 Kuz 和 TACE 的潜在 Notch 受体/配体底物在胸腺并分析 Notch 受体/配体的功能后果胸腺发育的分裂。我们预测在胸腺细胞中过度表达显性阴性 Kuz 或 TACE 基因，CD4+ T 细胞可能多于 CD8 T 细胞发展。相反，在过表达野生型 Kuz 或 TACE 的胸腺细胞中，更多 CD8+ T 细胞比 CD4+ T 细胞会发育。这些预测将受到检验体外和转基因小鼠中。已知Notch的表达和处理胸腺中的受体和配体将通过定量蛋白质印迹法进行评估印迹和通过流式细胞术评估 T 细胞发育。相关性这些系统中的处理和 T 细胞发育之间的关系将调查了。