The Role of Kuzbanian and TACE in T Cell Development

Kuzbanian 和 TACE 在 T 细胞发育中的作用

• 批准号: 6622373
• 负责人: JENNIFER O MANILAY
• 金额: $ 4.81万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 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/6622373
• 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

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途径 对决定细胞命运很重要我们实验室的研究首先 提出了Notch信号在胸腺发育中的作用，导致胸腺发育异常。 接受Notch信号的未成熟胸腺细胞发育为CD 8 + T细胞的模型 在一些情况下，不接受Notch信号的T细胞成熟为CD 4 + T细胞。 Notch-1受体的蛋白水解加工是产生Notch-1受体所必需的。 Notch-1介导的信号，而Delta，一种Notch配体， 在果蝇中加工。然而，小鼠Notch同源物的加工是否 和Notch配体发生，所有负责这种加工的蛋白酶 不知道。金属蛋白酶Kuzbanian（Kuz）和TACE已经被发现。 参与Notch受体和配体的加工。作为凹口的加工 是Notch信号传导所需的，是Notch配体加工的阻断剂， 受体可能导致Notch信号传导减少。我们将确定 表达的Kuz和TACE的潜在Notch受体/配体底物 胸腺和分析Notch受体/配体的功能后果 胸腺发育时的卵裂。我们预测，在胸腺细胞中， 显性阴性Kuz或TACE基因，CD 4 + T细胞多于CD 8 T细胞可能 开发.相反，在过度表达野生型Kuz或TACE的胸腺细胞中， CD 8 + T细胞比CD 4 + T细胞会发展。这些预测将得到检验 在体外和转基因小鼠中。已知Notch的表达和加工 胸腺中的受体和配体将通过定量Western印迹法进行评估。 印迹法和通过流式细胞术评估的T细胞发育。相关性 在这些系统中，加工和T细胞发育之间的关系将是 研究了