High throughput quantification of conformation and kinetics of DNA-protein complexes

DNA-蛋白质复合物的构象和动力学的高通量定量

• 批准号: 0933670
• 负责人: Selim Unlu
• 金额: $ 33万
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2014-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0933670&HistoricalAwards=false
• 关键词: throughput; quantification; conformation; kinetics; DNA

0933670Unlu This NSF award by the Biosensing /CBET program supports work by Professor Ünlü at Boston University to study conformation and kinetics of DNA-protein complexes on a high-throughput platform. DNA function and DNA repair depend on interactions with proteins - histones, transcriptional proteins and nucleases among them. The regulation of gene transcription involves the formation of specific protein - DNA complexes that bring distant regions of DNA together and can sharply bend or kink the DNA. These conformational changes are postulated to play an important role in the recognition of specific binding sites on DNA by proteins and thus detailed understanding of the conformation and dynamics of formation of specific complexes are of considerable biological significance. Yet current technology does not exist to access the length scales necessary to directly measure these biomolecular conformation. Current fluorescence methods do not provide the necessary resolution and while the electron microscopy has high resolution it destroys the sample. This research program will develop and demonstrate a platform technology for the precise quantified measurement of conformation of DNA-protein complexes in their native environment. Our approach combines innovations from Boston University (BU) and Technical University of Munich (TUM), merging optical interference techniques with surface electric field driven, highly-ordered DNA arrays. A method that precisely measures the shape of DNA-protein complexes in a high-throughput array format can address a range of important questions about the biophysics of the conformation and orientation of DNA-protein complexes. The success of this proposed program will provide a unique tool with significant cost reduction and increase in speed in structural measurements of DNA-protein complexes and allow for the study the basic principles of regulation and expression of genes. The significant educational outcome of this program will be training at least two graduate students. The students involved in the research will have the opportunity to practice and perfect communication skills within a professional context through journal clubs, summer schools, research seminars, and participation in local, national, and international conferences. They will work closely with international collaborators from Germany and Singapore. Students also will have the experience and training of expositing to a lay audience, best exemplified by presenting in middle and high school classrooms, or working with teachers in summer research and immersive science classes, or with urban high school students in lab research to build motivation to pursue a STEM career.

0933670unlu Biossysing /CBET计划的NSF奖支持波士顿大学的ünlü教授的工作，在高通量平台上为DNA-蛋白综合体的研究会议和动力学。 DNA功能和DNA修复取决于与蛋白质，组蛋白，转录蛋白和核的相互作用。基因转录的调节涉及将特定蛋白质 - DNA复合物的形成，这些蛋白DNA复合物将遥远的DNA区域聚集在一起，并可以急剧弯曲或扭结DNA。假定这些构象变化是通过蛋白质识别特定结合位点在DNA上的重要作用的重要作用，因此对特定复合物形成的组成和动力学的详细理解具有可考虑的生物学意义。然而，目前的技术并不存在直接测量这些生物分子构象所必需的长度尺度。当前的荧光方法没有提供必要的分辨率，而电子显微镜具有高分辨率，它会破坏样品。该研究计划将开发并展示一种平台技术，用于在其本地环境中精确量化DNA-蛋白质复合物。我们的方法结合了波士顿大学（BU）和慕尼黑技术大学（TUM）的创新，将光学干扰技术与表面电场驱动，高度有序的DNA阵列合并。一种精确测量高通量阵列格式DNA蛋白复合物形状的方法可以解决有关DNA-蛋白复合物组成和方向的生物物理学的一系列重要问题。该提出的程序的成功将为DNA-蛋白质复合物的结构测量速度大幅降低和速度提高提供独特的工具，并允许研究基因的调节和表达的基本原理。该计划的重大教育结果将是培训至少两名研究生。参与研究的学生将有机会通过期刊俱乐部，暑期学校，研究半读物以及参加当地，国家和国际会议，在专业背景下练习和完美的沟通技巧。他们将与来自德国和新加坡的国际合作者紧密合作。学生还将获得与外行受众接触的经验和培训，最好通过在中学和高中教室中展示，或与夏季研究和身临其境科学课程的老师合作，​​或与实验室研究中的城市高中生一起工作以建立从事STEM职业的动力。