MRPG: In vivo Characterization of the Role of eIF 2B epsilonPhosphorylation in Control of Protein Synthesis

技术摘要：这项少数民族研究计划拨款旨在为一个完整的提案奠定基础，该提案将专注于eIF-2B在真核细胞中蛋白质合成调节中的作用。具体来说，将开发一个哺乳动物表达系统来研究eIF-2B最大亚基的磷酸化在控制蛋白质合成中的作用。将该亚基cDNA克隆并转染到COS-7细胞中。目前手头的cDNA是不完整的，全长cDNA将使用限制性内切酶构建。一个2-D凝胶电泳系统将被开发，将允许表征的磷酸化状态的eIF-2B。将为竞争性赠款提案编制初步数据。摘要：高等动物和植物的蛋白质合成是由一组称为起始因子的蛋白质调节的。本项目侧重于表征一个起始因子，eIF-2 α。在病毒感染、重金属暴露、营养剥夺和热休克等环境胁迫下，EIF-2 α介导蛋白质合成的抑制。这项研究可以更好地理解压力下蛋白质合成的调节。

Asuru 9724040 Technical Abstract: This minority research planning grant is aimed at laying the groundwork for a full proposal that would focus on the role of eIF-2B in the regulation of protein synthesis in eukaryotic cells. Specifically, a mammalian expression system will be developed to study the role of phosphorylation of the largest subunit of eIF-2B in controlling protein synthesis. The cDNA for this subunit will be cloned and transfected into COS-7 cells. The cDNA currently in hand is incomplete, and the full length cDNA will be constructed using restriction enzymes. A 2-D gel electrophoresis system will be developed that will allow the characterization of the phosphorylation states of eIF-2B. Preliminary data will be developed for a competitive grant proposal. Non-Technical Abstract: Protein synthesis in higher animals and plants is regulated by a group of proteins called initiation factors. This project focuses on characterizing one initiation factor, eIF-2 alpha. EIF-2 alpha mediates the inhibition of protein synthesis in response to environmental stresses such as viral infection, heavy metal exposure, nutrient deprivation and heat shock. This research could lead to a better understanding of regulation of protein synthesis during stress.