DNA-Protein Interactions Studied Directly Using Single Molecule Fluorescence Imaging of Quantum Dot Tagged Proteins Moving on DNA Tightropes.
Many protein interactions with DNA require specific sequences; however, how these sequences are located remains uncertain. DNA normally appears bundled in solution but, to study DNA-protein interactions, the DNA needs to be elongated. Using fluidics single DNA strands can be efficiently and rapidly elongated between beads immobilized on a microscope slide surface. Such "DNA tightropes" offer a valuable method to study protein search mechanisms. Real-time fluorescence imaging of these interactions provides quantitative descriptions of search mechanism at the single molecule level. In our lab, we use this method to study the complex process of nucleotide excision DNA repair to determine mechanisms of damage detection, lesion removal, and DNA excision.
School of Biosciences, University of Kent, Canterbury, CT2 7NH, UK.
Luke
L
Springall
School of Biosciences, University of Kent, Canterbury, CT2 7NH, UK.
Alessio V
AV
Inchingolo
School of Biosciences, University of Kent, Canterbury, CT2 7NH, UK. n.kad@kent.ac.uk.
Neil M
NM
Kad
D000465:Algorithms; D001665:Binding Sites; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D044085:Microfluidics; D008856:Microscopy, Fluorescence; D036103:Nanotechnology; D011485:Protein Binding; D045663:Quantum Dots; D000072760:Single Molecule Imaging / Q000379:methods*
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins
DNA repair; DNA tightropes; Diffusion; Nucleotide excision repair; Quantum dots; Search mechanisms; Single molecule imaging