Fluorescence Fluctuation Spectroscopy to Investigate Molecular Recognition Specificity

该项目的目的是开发先进的单分子光谱方法，用于定量分析蛋白质与核酸的相互作用，并在其应用中了解锌指蛋白Nab2以序列特异性与核酸结合并区分单链RNA和DNA的物理机制。蛋白质-核酸相互作用控制着基因表达的所有阶段，从基因组的维持到RNA表达，再到翻译和蛋白质合成。该项目有两个广泛的研究目标：推进荧光波动光谱（FFS）方法作为研究分子相互作用的定量工具；并利用这些方法研究了一种重要的mRNA加工和输出蛋白——Saccharomyces cerevisiae Nuclear Abundant Poly(A) Binding protein （Nab2）与核酸相互作用中分子识别的生物物理机制。这些方法将广泛应用于分子相互作用的研究。这里描述的项目是Berland（埃默里学院物理系）和Corbett（埃默里医学院生物化学系）实验室的合作成果。该项目将通过教学和培训机会产生广泛的影响，使本科生和研究生能够在物理和生物化学的界面上工作。重要的是，这些研究将为实验室和课堂上的跨学科学习奠定基础。这项工作还提供了与埃默里大学以外的社区交流的机会，因为两个pi实验室都有与亚特兰大高中互动的历史，通过在实验室接待学生和教师。

The objective of this project is to develop advanced single molecule spectroscopy methods for quantitative analysis of protein-nucleic acid interactions, as well as in their application to understand the physical mechanisms by which the zinc finger protein Nab2 binds to nucleic acids with sequence specificity and discriminates between single stranded RNA and DNA. Protein-nucleic acid interactions control all phases of gene expression from maintenance of the genome to RNA expression to translation and protein synthesis. This project has two broad research aims: advancing fluorescence fluctuation spectroscopy (FFS) methods as quantitative tools to investigate molecular interactions; and applying these methods to investigate the biophysical mechanisms underlying molecular recognition in the interaction of an essential mRNA processing and export protein, Saccharomyces cerevisiae Nuclear Abundant Poly(A) Binding Protein (Nab2) with nucleic acids. The methods developed will be widely useful for molecular interaction studies. The projects described here are the collaborative effort of the Berland (Physics Department, Emory College) and Corbett (Biochemistry Department, Emory School of Medicine) laboratories. This project will have a broad impact through teaching and training opportunities that enable undergraduate and graduate students to work at the interface between physics and biochemistry. Importantly, these studies will set the stage for interdisciplinary learning both in the laboratory and in the classroom. This work also affords opportunities for interfacing with the community beyond Emory as both PIs laboratories have a history of interactions with high schools in Atlanta through hosting both students and teachers in the lab.