Regulatory Protein-DNA Interactions in vivo Analyzed by Ultrafast Photochemical Crosslinking

瓦特W.应用与工程物理学院，约翰T。分子生物学和遗传学系，化学和化学生物学系，Bik-Kwoon Tye，分子生物学和遗传学系，Eric E。分子生物学和遗传学系的Alani和亚历山大L.盖塔，应用和工程物理学院，康奈尔大学所有的资助与化学计划的合作研究奖，由化学部，物理部，分子，细胞生物学部和多学科活动办公室提供的资金。 这个多方面的团队将采用超快紫外激光交联技术，在细胞周期中发生功能激活时，将蛋白质瞬间共价锁定在其核酸靶点上。然后通过最先进的分析蛋白质和核酸生物化学和结构分析来解析这些分子复合物，以鉴定交联的氨基酸及其核苷酸结合位点。这些研究人员将研究复杂的DNA-蛋白质相互作用的网络，这些网络调节基因转录、复制和修复，使用新技术可以对活细胞中难以捉摸的DNA-蛋白质相互作用进行高时空分辨率的生化分析。 本研究的最终目标是发展在原核和真核系统活细胞上进行功能基因组实验的方法，以建立DNA结合蛋白的全基因组图谱，从而跟踪整个细胞生长周期的基因调控，并为破译基因调控网络的复杂性提供有效的工具。将采用协作方法来靶向DNA结合蛋白的光化学和功能基因组学。科学思想将跨三个学科-生物学，化学和物理学-联合分子生物学家研究DNA-蛋白质相互作用;化学家研究DNA的光化学;物理学家应用超短激光脉冲光学;生物物理学家为跨学科研究提供接口。学生将接受从高中到博士后的各级培训。

Watt W. Webb, School of Applied and Engineering Physics, John T. Lis, Department of Molecular Biology and Genetics, Tadhg P. Begley, Department of Chemistry and Chemical Biology, Bik-Kwoon Tye, Department of Molecular Biology and Genetics, Eric E. Alani, Department of Molecular Biology and Genetics, and Alexander L. Gaeta, School of Applied and Engineering Physics, all of Cornell University are funded with an award from the Collaborative Research in Chemistry Program with funds provided by the Division of Chemistry, the Division of Physics, the Division of Molecular, Cellular Biology and the Office of Multidisciplinary Activities. This multifaceted team will employ ultrafast UV laser crosslinking to lock proteins to their nucleic acid targets instantaneously and covalently during functional activation as it occurs during the cell cycle. These molecular complexes are then resolved by state-of-the-art analytical protein and nucleic acid biochemistry and structural analysis to identify crosslinked amino acids and their nucleotide binding-sites. These researchers will investigate the network of complex DNA-protein interactions regulating gene transcription, replication, and repair using new technologies that allow high temporal and spatial resolution biochemical analysis of elusive DNA-protein interactions in living cells. The ultimate goal of this research is to develop the methods to perform functional genomic experiments on living cells of prokaryotic and eukaryotic systems to establish genome-wide maps of DNA-bound proteins, thus following global gene regulation control through a cell's growth cycle and providing efficient tools for deciphering the complexity of gene regulator networks.In this research, a collaborative approach will be employed to target both the photochemistry and the functional genomics of DNA-binding-proteins. Scientific ideas will be integrated across three disciplines - biology, chemistry, and physics - uniting molecular biologists investigating DNA-protein interactions; chemists studying the photochemistry of DNA; physicists applying ultrashort laser pulse optics; and biophysicists providing the interface for the interdisciplinary research. Students will receive training at all levels from high school to postdoctoral.