SLAP 130 AND THE REGULATION OF T CELL ACTIVATION

SLAP 130 和 T 细胞激活的调节

• 批准号: 2709458
• 负责人: NANCY J BOERTH
• 金额: $ 3.02万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-11-30 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2709458
• 关键词: CD antigens; T cell receptor; T lymphocyte; binding proteins; biological signal transduction; calcium flux; cell differentiation; chimeric proteins; enzyme activity; flow cytometry; gene expression; gene targeting; genetically modified animals; immunofluorescence technique; inositol phosphates; laboratory mouse; leukocyte activation /transformation; mitogen activated protein kinase; phosphorylation; protein structure function; protein tyrosine kinase; site directed mutagenesis; thymus

Engagement of the T-cell antigen receptor (TCR) results in protein tyrosine kinase (PTK) activation followed by the subsequent phosphorylation of numerous proteins. Previous studies which focused on identifying substrates of TCR-induced PTK activity led to the cloning and characterization of SLP76. In sight into the function of SLP76 WAS provided by the findings that SLP76 regulates TCR-mediated signals that leads to induction of the IL-2 gene promoter. In order to investigate further the function of SLP76 in T-cell activation, we and others have begun to characterize SLP-76 associated proteins. In this study, we focus our attention on the recently identified SLP76 associated phosphoprotein of 130kDa, SLAP130. Sequence analysis reveals that SLAP130 contains several regions that may mediate its interaction with other molecules although it lacks any known enzymatic activity. Initial experiments show that overexpression of SLAP130 interferes with TCR signaling and can inhibit the ability of SLP76 to augment NFAT activity. Although the mechanism of how SLAP130 modulates T-cell activation remains unclear, our preliminary studies provide support for the hypothesis that SLAP130 may serve as a negative regulator of T-cell activation. To test this hypothesis, initial experiments will involve the characterization of the expression pattern of SLAP130 in hematopoietic tissues and during development and T-cell maturation. In addition, we will investigate which proximal TCR-mediated signals are affected by SLAP130. The next set of experiments will focus on understanding the structure/function relationship of SLAP130. We will identify the tyrosine phosphorylation sites in SLAP130 and will ask which of these are responsible for the interaction between SLAP130 and SLP76. In order to understand better the function of SLAP130, we propose to identify other proteins that associate with SLAP130 and will ask whether these interactions are important in modulating T-cell function. Finally, to address the importance of SLAP130 function in T-cells, we will use a SLAP130 "knock-out" mouse to assess T-cell function in the absence of SLAP130 expression. It is hoped that the information gained from these studies will provide insight into the function of SLAP130 in modulating T-cell signaling.

T 细胞抗原受体 (TCR) 的参与会产生蛋白质 酪氨酸激酶（PTK）激活，随后 许多蛋白质的磷酸化。先前的研究主要集中于 鉴定 TCR 诱导的 PTK 活性底物导致克隆 和 SLP76 的表征。深入了解SLP76 WAS的功能 SLP76 调节 TCR 介导的信号这一发现提供了 导致 IL-2 基因启动子的诱导。为了调查 我们和其他人进一步研究了 SLP76 在 T 细胞激活中的功能 开始表征 SLP-76 相关蛋白。在这项研究中，我们 将我们的注意力集中在最近发现的 SLP76 相关 130kDa 的磷蛋白，SLAP130。序列分析表明 SLAP130 包含几个可能介导其相互作用的区域 尽管它缺乏任何已知的酶活性，但它与其他分子无关。最初的 实验表明SLAP130的过度表达会干扰TCR 信号传导并可以抑制 SLP76 增强 NFAT 活性的能力。 尽管 SLAP130 调节 T 细胞激活的机制 仍不清楚，我们的初步研究为 假设 SLAP130 可能作为 T 细胞的负调节因子 激活。为了检验这个假设，最初的实验将涉及 SLAP130 表达模式的表征 造血组织以及发育和 T 细胞成熟过程中。在 此外，我们将研究哪些近端 TCR 介导的信号是 受 SLAP130 影响。下一组实验将集中于 了解 SLAP130 的结构/功能关系。我们将 识别 SLAP130 中的酪氨酸磷酸化位点并询问 其中哪些负责 SLAP130 和 SLAP130 之间的相互作用 SLP76。为了更好地理解SLAP130的功能，我们建议 识别与 SLAP130 相关的其他蛋白质，并询问 这些相互作用对于调节 T 细胞功能是否重要。 最后，为了解决 SLAP130 功能在 T 细胞中的重要性，我们 将使用 SLAP130“敲除”小鼠来评估 T 细胞功能 SLAP130 表达缺失。希望所获得的信息 这些研究将深入了解 SLAP130 在 调节 T 细胞信号传导。