TRANSCRIPTIONAL REGULATION BY THE CAM CASCADE

CAM 级联的转录调控

• 批准号: 6105486
• 负责人: THOMAS R SODERLING
• 金额: $ 19.47万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6105486
• 关键词: PC12 cells; biological signal transduction; cAMP response element binding protein; calcium flux; calmodulin dependent protein kinase; cell line; enzyme inhibitors; enzyme mechanism; gene expression; genetic regulatory element; genetic transcription; mitogen activated protein kinase

Elevated intracellular calcium is one of the most important signaling molecules in cells, especially in neural tissues. One of the important actions of calcium is to regulate the transcription of selected genes, thereby altering the phenotype of the cell. Studies from a number of laboratories, including our own, have demonstrated that a family of calmodulin-dependent protein kinases mediate many of these Ca2+- dependent transcriptional events. In particular, we have focused on CaM-kinase IV which can phosphorylate several transactivating proteins such as CREB and SRF. In the past two years we have partially characterized a CaM-kinase cascade consisting of a CaM- kinase kinase which can phosphorylate and activate CaM-kinase IV and CaM-kinase I. In the current grant application we will further characterize the involvement of this CaM-kinase cascade by: 1) identifying additional upstream components of the cascade (e.g. CaM- kinase kinase kinase) or anchoring proteins for CaM-kinase kinase. 2) identifying additional transactivating proteins for CaM-kinase IV by examining transcriptional regulation of the NGFI-B gene by the CaM- kinase cascade. 3) determining the sites and physiological relevance of cross-talk between the CaM-kinase cascade and the MAP-kinase cascade. These studies will be performed in a variety of cultured cells, especially PC12 cells, and will utilize biochemical and molecular biological techniques. The results of this study have implications for modulating cellular apoptosis and synaptic plasticity due to Ca2+- dependent gene transcription.

升高细胞内钙是最重要的信号之一 细胞中的分子，尤其是在神经组织中。 重要的之一 钙的作用是调节所选基因的转录， 从而改变细胞的表型。 来自多个的研究 包括我们自己在内的实验室已经证明了一个家庭 钙调蛋白依赖性蛋白激酶介导了许多CA2+ - 依赖的转录事件。 特别是，我们专注于 CAM激酶IV可以磷酸化几个反式激活 蛋白质，例如CREB和SRF。 在过去的两年中 部分表征了由凸轮组成的凸轮激酶级联 激酶激酶可以磷酸化并激活Cam-激酶IV，并且 凸轮激酶I。在当前的赠款申请中，我们将进一步 表征该凸轮激酶级联的参与：1） 识别级联的其他上游组成部分（例如 激酶激酶激酶）或用于CAM-激酶激酶的锚定蛋白。 2） 通过通过 检查CAM-对NGFI-B基因的转录调控 激酶级联。 3）确定站点和生理相关性 凸轮激酶级联和地图激酶之间的串扰 级联。 这些研究将在多种培养的细胞中进行， 特别是PC12细胞，并将使用生化和分子 生物技术。 这项研究的结果对 根据Ca2+ - 调节细胞凋亡和突触可塑性 依赖基因转录。