Molecular dissection of eukaryotic transcription termination mechanisms

对于真核细胞来说，一个巨大的挑战是生产适量的每种蛋白质，并随着细胞环境的变化迅速调整产量。为了迎接这一挑战，细胞必须协调许多离散的步骤，但这种整合的细节还没有被很好地理解。这项研究将以面包酵母为模型，探索细胞用来对环境压力做出综合反应的分子相互作用。因为所有生物体都必须应对各种类型的压力，所以结果应该适用于其他系统，并可能有助于制定工程压力耐受性策略。除了增进科学知识外，该项目还将为来自不同背景的本科生、毕业后实习生和研究生提供培训机会。它将通过为主要来自本科院校的初级教师在其职业生涯的关键时刻提供研究机会来改善STEM教育。该项目还将提供计算科学方面的培训，作为生物学研究的定量和预测工具，并开发一个公开可用的网络界面，供研究人员使用全基因组数据集探索基因调控的各个方面。对细胞压力的适当反应需要许多基因表达步骤之间的协调。这些步骤包括信使核糖核酸合成的不同阶段--启动、延长和终止--以及加工过程中的多个步骤，如封端、剪接、聚腺苷酸化，所有这些都以从细胞核输出信使核糖核酸为最终结果。这个“协调网络”的每个部分都构成了一个步骤，在这个步骤中，来自特定基因的信使核糖核酸的数量和类型可以被调节。该项目的目标是使用酵母酿酒酵母作为模式生物来确定协调转录和mRNA3‘末端加工的关键分子相互作用。特别是，该项目将使用分子生物学、生物化学、遗传学和生物信息学的互补方法来确定核心多聚腺苷化因子如何在应激反应中对基因的激活和抑制做出贡献。这些结果应该会为细胞用来快速微调其对不断变化的环境条件的反应的机制提供新的见解。

A great challenge for a eukaryotic cell is to produce the appropriate amount of each protein and to quickly adjust that production as the cell's environment changes. To meet this challenge, cells must orchestrate many discrete steps, but the details of this integration are not well understood. This research will use baker's yeast as a model to explore the molecular interactions cells use to mount an integrated response to environmental stress. Because all organisms must cope with stresses of various types, the results should be applicable to other systems and may aid in developing strategies for engineering stress tolerance. In addition to advancing scientific knowledge, this project will provide opportunities for training undergraduates, post-baccalaureate trainees, and graduate students from diverse backgrounds. It will improve STEM education by providing research opportunities for junior faculty from primarily undergraduate institutions at critical points in their careers. The project will also provide training in computational science as a quantitative and predictive tool for biological studies, and develop a publicly available web interface for researchers to explore aspects of gene regulation using whole genome datasets. Proper responses to cellular stress require coordination among a number of gene expression steps. These include different phases of mRNA synthesis--initiation, elongation and termination--as well as multiple steps in processing, such as capping, splicing, polyadenylation, all culminating with export of the mRNA from the nucleus. Each part of this "coordination network" constitutes a step at which the amount and type of mRNA from a particular gene can be regulated. The goal of this project is to use the yeast Saccharomyces cerevisiae as a model organism to identify critical molecular interactions that coordinate transcription and mRNA 3' end processing. In particular, the project will use complementary approaches of molecular biology, biochemistry, genetics, and bioinformatics to determine how core polyadenylation factors contribute to the activation and repression of genes in response to stress. The results should provide new insights into the mechanisms cells use to rapidly fine-tune their responses to changing environmental conditions.