Collaborative Research - CPSF30 at the convergence of cellular signaling and RNA Processing

非技术总结：环境压力（来自干旱和高温或低温等条件）是作物生产力的重要和持续关注的问题。包括农作物在内的植物会以多种方式对来自环境的信号做出反应。这些反应包括基因表达方式的变化。了解和控制这些变化可能是提高作物生产力的有利手段。本研究的主题是一种蛋白质（称为AtCPSF30），它对植物对环境胁迫的反应以及指导从幼苗到成熟和生产植物的其他线索（或发育信号）非常重要。这项研究的目的是了解这种蛋白质如何帮助植物区分不同的信号，从而使随后的基因表达变化与各自的压力或发育信号相适应。这项研究将有助于更好地了解植物如何应对环境胁迫，以及胁迫如何影响生长发育的各个方面。这种理解将使科学家能够开发改良的作物品种，并将帮助生产者在作物和品种选择以及生产管理方面做出明智的决定。项目技术描述：本研究的主题是一个假想的中枢，它将细胞信号转导成差异多聚(a)位点选择和表型结果。这个中心是所谓的拟南芥裂解和聚腺苷酸化特异性因子（AtCPSF30）的30 kD亚基，在体外受两种不同的化学或生化信号（钙和氧化还原信号）调节，并参与一系列发育和生理反应。本研究的目的是确定这些不同的信号线索是如何转导到特定的结果的，可能是通过mrna前加工过程中的选择性聚腺苷酸化（APA）。目标将通过测试这三个假设来实现：1。不同的信号输入通过AtCPSF30被处理成不同的APA事件和结果。3. atcpsf30介导的APA可改变mRNA的功能，从而影响该蛋白的一些直接靶点。atcpsf30介导的APA影响侧根发育早期的基因表达。将广泛使用系统方法——高通量DNA测序和大数据集的计算分析。这些研究将由基因、转基因、细胞、分子和生化研究补充，所有这些研究都将用于上述假设的检验。这项研究的结果将进一步了解RNA加工如何将信号信号整合到不同的分子结果中，以及这些结果如何影响遗传网络，进而影响基本的生长过程。这个项目的数据将酌情纳入课堂，让学生了解研究过程如何运作，以及如何收集和分析大规模数据集的第一手信息。在提议的研究过程中产生的实验室和计算专业知识以及数据集将提供给一个学院和大学联盟，该联盟的目标是促进实验室和课堂练习的发展，旨在使本科生熟悉高通量DNA测序。其他拓展活动包括：与地区HBCU和高中的教师合作，以及（由李博士）参与一个旨在招募和指导少数民族学生进行细胞和分子生物学研究的项目。

Non-technical summary:Environmental stresses (from conditions such as drought and high or low temperatures) are significant and ongoing concerns for crop productivity. Plants, including crops, respond to cues from the environment in many ways. Among these responses are changes in the ways that genes are expressed. Understanding and manipulating these changes are likely to be profitable means for improving crop productivity. The subject of this research is a protein (termed AtCPSF30) that is important for the responses of plants to environmental stresses, as well as to other cues (or developmental signals) that guide development from a seedling to a mature and productive plant. The goals of the research are to understand how this protein helps the plant to discriminate between different cues, so that subsequent changes in gene expression are appropriate to the respective stress or developmental signal. This research will lead to a better understanding of how plants respond to environmental stresses, and to how stresses affect different aspects of growth and development. This understanding will allow scientists to develop improved crop varieties, and will help producers to make informed decisions regarding crop and variety choice, as well as production management. Technical description of the project:The subject of this study is a hypothetical hub that transduces cellular signaling cues into differential poly(A) site choices and phenotypic outcomes. This hub, the 30 kD subunit of the so-called Cleavage and Polyadenylation Specificity Factor in Arabidopsis (AtCPSF30), is regulated in vitro by two distinctive chemical or biochemical signals (calcium and redox signals), and is involved in a range of developmental and physiological responses. The goals of the research are to determine how these different signaling cues are transduced to specific outcomes, potentially through alternative polyadenylation (APA) during pre-mRNA processing. The goals will be accomplished by testing these three hypotheses:1. Different signaling inputs are processed, through AtCPSF30, into differing sets of APA events and outcomes.2. AtCPSF30-mediated APA alters mRNA functionality for some of the numerous direct targets of the protein.3. AtCPSF30-mediated APA affects gene expression early in lateral root development.Extensive use will be made of systems approaches - both high-throughput DNA sequencing and computational analyses of large datasets. These will be complemented by genetic, transgenic, cellular, molecular, and biochemical studies, all of which will be brought to bear on tests of the stated hypotheses. The outcomes of this research will further an understanding of how RNA processing integrates signaling cues into distinctive molecular outcomes, and how these outcomes affect genetic networks that in turn impact essential growth processes. The data from this project will be incorporated as appropriate into classes to give the students first-hand information on how the research process works, and how large-scale datasets are collected and analyzed. The laboratory and computational expertise and the datasets that will be generated in the course of the proposed studies will be made available to a consortium of colleges and universities that has the goal of facilitating the development of laboratory and classroom exercises intended to familiarize undergraduate students with high-throughput DNA sequencing. Other outreach activities include: a collaboration with faculty at regional HBCU and highs schools and participation (by Dr. Li) in a project intended to recruit and mentor minority students in research in cell and molecular biology.