NSF/MCB-BSF: Integrating ROS, redox and cell metabolism across plant and animal cells

自从大约27亿年前光合作用生物将氧气引入大气以来，活化形式的氧(称为活性氧)一直是有氧生命中不受欢迎的伴侣。虽然目前被植物和动物细胞作为重要的信号分子使用，但这些被激活的氧形式对细胞和组织可能具有高度毒性，并导致氧化损伤(氧化应激)。该项目由北得克萨斯大学的Ron Mittler和Rajeev Azad以及以色列耶路撒冷希伯来大学的Rachel Nechushtai领导，其长期目标是确定细胞如何监控细胞内的活性氧水平，并防止其毒性。特别是，该项目将突出植物和动物细胞中活性氧调节的一个未知方面，即一组新发现的蛋白质利用铁-硫簇来监测活性氧水平，并调节细胞新陈代谢和其他重要过程。这项研究的结果可能导致开发新的和新的方法来提高作物对干旱和高温等重要逆境的耐受性或延缓衰老。此外，这项拟议的研究还可以确定新的植物性化合物和蛋白质，以缓解氧化应激、衰老和不同的疾病，如癌症和糖尿病。PIS将培训一些研究生和本科生，并与当地的教育中心和博物馆合作，为K-12学生提供外展服务。PIS将研究一类新的Fe-S蛋白质-NEET蛋白质在维持动植物细胞ROS动态平衡中的作用。根据其独特的簇特征，假设NEET蛋白使用其氧化还原活性不稳定的簇来感知细胞中的ROS水平，并调节改变细胞代谢的不同途径。该项目的具体目标是：1.对NEET蛋白水平和/或功能改变的动植物细胞进行比较信号和调控网络分析。2.鉴定动植物细胞的NEET相互作用组网络。3.确定NEET蛋白在细胞中定位/功能的动态，并进行哺乳动物和植物细胞之间的NEET蛋白的遗传互补研究。结合功能遗传学、蛋白质组学、先进成像、RNA-Seq和网络分析方法，将寻求对细胞中ROS/氧化还原反应/调节的机械性理解。通过这项拟议的NSF-BSF合作确定的机制将在不同的王国之间进行进一步的比较：通过NSF部分研究植物，通过BSF部分研究动物，以获得细胞中ROS/氧化还原感应/调节的进化观点。这项美国和以色列的合作项目得到了美国国家科学基金会和以色列双国科学基金会的支持。

Ever since the introduction of oxygen into the atmosphere by photosynthetic organisms, about 2.7 billion years ago, activated forms of oxygen (called reactive oxygen) have been the unwelcome companions of aerobic life. Although currently used by plant and animal cells as important signaling molecules, these activated forms of oxygen could be highly toxic to cells and tissues and cause oxidative injury (oxidative stress). The long-term goal of this project, led by Ron Mittler and Rajeev Azad of the University of North Texas and Rachel Nechushtai of Hebrew University in Jerusalem, Israel, is to determine how cells monitor their intracellular levels of reactive oxygen and prevent its toxicity. In particular, the project will highlight an unknown aspect of the regulation of reactive oxygen in plant and animal cells, namely the use of iron-sulfur clusters by a newly discovered group of proteins to monitor reactive oxygen levels and regulate cellular metabolism and other vital processes. Results obtained from this study could lead to the development of new and novel approaches to enhance the tolerance of crops to important stresses such as drought and heat or delay senescence. In addition, the proposed study could identify novel plant-based compounds and proteins that mitigate oxidative stress, aging and different diseases such as cancer and diabetes. The PIs will train a number of graduate and undergraduate students and partner with a local education center and museum to provide outreach to K-12 students.The PIs will investigate the role of a novel class of Fe-S proteins, NEET proteins, in maintaining ROS homeostasis in plant and animal cells. In light of their unique cluster features, it is hypothesized that NEET proteins use their redox-active labile clusters to sense ROS levels in cells and regulate different pathways that alter cellular metabolism. The Specific Aims of the project are: 1. Perform a comparative signaling and regulatory network analysis of plant and animal cells with altered level and/or function of NEET proteins. 2. Identify the NEET interactome network of plant and animal cells. 3. Determine the dynamics of NEET protein localization/ function in cells and conduct genetic complementation studies of NEET proteins between mammalian and plant cells. Using a combination of functional genetics, proteomics, advanced imaging, RNA-Seq and network analysis approaches, a mechanistic understanding of ROS/redox sensing/regulation in cells will be pursued. The mechanisms identified through this proposed NSF-BSF collaboration will be further compared between different kingdoms: plant - studied through the NSF part, and animal - studied through the BSF part of this project, to obtain an evolutionary perspective of ROS/redox sensing/regulation in cells.This collaborative US/Israel project is supported by the US National Science Foundation and the Israeli Binational Science Foundation.