NOVEL T CELL ACTIVATION GENE IN DEVELOPING NEURONS

发育中神经元中的新型 T 细胞激活基因

• 批准号: 2445529
• 负责人: MICHAEL B PRYSTOWSKY
• 金额: $ 30.03万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2445529
• 关键词: DNA binding protein; T lymphocyte; behavioral /social science research tag; gene expression; gene mutation; genetic regulatory element; genetically modified animals; interleukin 2; laboratory mouse; laboratory rabbit; leukocyte activation /transformation; neurogenesis; phosphorylation; potassium channel; psychoneuroimmunology; tissue /cell culture

DESCRIPTION (Adapted from applicant's abstract): Mental effects on physical health are mediated by interactions between the nervous and immune systems. These systems are coordinately regulated through common molecules. Given the common expression of cell surface receptors and soluble mediators in both systems, one might expect to find intracellular proteins that are common to both systems and may be involved in signaling. The investigators isolated a cDNA sequence from a cDNA library prepared from IL2-stimulated, cloned T lymphocytes which has recently been identified as the KvBeta2 subunit of voltage-gated K channels. Potassium channels maintain membrane potential and determine the excitability of cells. Beta subunits appear to regulate the conductance properties of K channels. KvBeta2 is expressed in proliferating lymphocytes and postnatal neurons. The hypothesis proposed is that Beta subunit expression affects lymphocyte and neuronal function through modulation of K channel conductance. The first aim will test the hypothesis that Beta subunit expression is required for IL2-driven T cell proliferation. The investigators indicate that with their knowledge of IL-2 induced gene expression, the precise point at which cell cycle progression is inhibited will be determined. In a second aim, studies will be initiated to define cis elements in a Beta subunit promoter and corresponding DNA binding proteins which are responsible for IL2-induced Beta subunit expression in lymphocytes. In a third aim, activation-dependent phosphorylation sites on Beta subunits will be determined in vitro and in vivo. Once specific phosphoamino acids are found, mutant proteins precluding specific phosphorylation will be prepared to determine the requirement of phosphorylation for Beta subunit function during IL2-driven proliferation. To determine if Beta subunit is required for lymphocytic and neuronal development and function, mice homozygous for a Beta subunit null allele will be prepared. While these animals may be immunodeficient at birth, the effect of Beta subunit deletion on neuronal development should not appear until postnatal neuronal maturation occurs. These studies will define the functional significance for KvBeta subunits in both the nervous and immune systems and begin to address molecular mechanisms regulating tissue specific expression.

描述（改编自申请人的摘要）：精神对身体的影响 健康是由神经系统和免疫系统之间的相互作用介导的。 这些系统通过共同的分子协调调节。 给定 细胞表面受体和可溶性介质的共同表达， 在这两种系统中，人们可能会期望找到细胞内的蛋白质， 对于两个系统是共同的，并且可以涉及信令。 调查人员 从IL 2刺激的cDNA文库中分离cDNA序列， 克隆的T淋巴细胞最近被鉴定为KvBeta2 电压门控K通道亚基。 钾通道维持膜 潜力和决定细胞的兴奋性。 β亚基似乎 调节K通道的电导特性。 KvBeta2表达于 增殖淋巴细胞和出生后神经元。 提出的假设是 β亚基的表达影响淋巴细胞和神经元的功能 通过调节K通道电导。 第一个目标将考验 IL 2驱动T细胞需要β亚基表达的假设 增殖 研究人员指出，由于他们对IL-2的了解， 诱导基因表达，细胞周期进程的精确点 将被禁止。 在第二个目标中， 为了定义β亚基启动子和相应DNA中的顺式元件， 负责IL 2诱导的β亚单位的结合蛋白 在淋巴细胞中的表达。 在第三个目标中，激活依赖性 将在体外和体内测定β亚基上的磷酸化位点。 vivo. 一旦发现特定的磷酸氨基酸， 将制备排除特异性磷酸化的蛋白质，以确定 在IL 2驱动过程中β亚基功能的磷酸化要求 增殖 为了确定β亚单位是否是淋巴细胞和 β亚基缺失纯合子小鼠的神经元发育和功能 将制备等位基因。 虽然这些动物可能是免疫缺陷， 出生后，β亚基缺失对神经元发育的影响应该 直到出生后神经元成熟才出现。 这些研究将 定义KvBeta亚基在神经和神经系统中的功能意义， 和免疫系统，并开始解决分子机制调节 组织特异性表达