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

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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.