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

自从大约27亿年前光合生物将氧气引入大气以来，活性氧（称为活性氧）一直是需氧生物不受欢迎的伙伴。虽然目前被植物和动物细胞用作重要的信号分子，但这些活化形式的氧可能对细胞和组织具有高毒性，并引起氧化损伤（氧化应激）。该项目由北德克萨斯大学的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测其细胞内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未知方面，即新发现的一组蛋白质使用铁硫簇来监测活性氧水平并调节细胞代谢和其他重要过程。这项研究的结果可能会导致新的方法的发展，以提高作物对干旱和高温等重要胁迫的耐受性或延缓衰老。此外，这项拟议的研究可以识别出新的植物化合物和蛋白质，它们可以减轻氧化应激、衰老和癌症和糖尿病等不同疾病。pi将培训一些研究生和本科生，并与当地教育中心和博物馆合作，为K-12学生提供外展服务。pi将研究一类新的Fe-S蛋白NEET蛋白在维持植物和动物细胞中ROS稳态中的作用。鉴于其独特的簇特征，我们假设NEET蛋白利用其氧化还原活性的不稳定簇来感知细胞中的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/或功能改变的植物和动物细胞进行信号传导和调控网络的比较分析。2. 确定植物和动物细胞的NEET相互作用网络。3. 确定NEET蛋白在细胞中的定位/功能动态，并在哺乳动物和植物细胞之间进行NEET蛋白的遗传互补研究。结合功能遗传学，蛋白质组学，高级成像，RNA-Seq和网络分析方法，将追求细胞中ROS/氧化还原传感/调节的机制理解。通过NSF-BSF合作确定的机制将进一步在不同领域之间进行比较：通过该项目的NSF部分研究植物，通过BSF部分研究动物，以获得细胞中ROS/氧化还原传感/调控的进化视角。这个美国/以色列合作项目由美国国家科学基金会和以色列两国科学基金会支持。

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.