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Lee, Ja Yil
School of Life Sciences
Research Interests
  • Investigated eukaryotic homologous recombination.

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Measuring intermolecular rupture forces with a combined TIRF-optical trap microscope and DNA curtains

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Title
Measuring intermolecular rupture forces with a combined TIRF-optical trap microscope and DNA curtains
Author
Lee, Ja YilWang, FengFazio, TeresaWind, ShalomGreene, Eric C.
Keywords
Combination;  DNA curtains;  DNA pulling;  Optical trap;  Rupture force;  Single-molecule
Issue Date
2012-10
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Citation
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, v.426, no.4, pp.565 - 570
Abstract
We report a new approach to probing DNA-protein interactions by combining optical tweezers with a high-throughput DNA curtains technique. Here we determine the forces required to remove the individual lipid-anchored DNA molecules from the bilayer. We demonstrate that DNA anchored to the bilayer through a single biotin-streptavidin linkage withstands similar to 20 pN before being pulled free from the bilayer, whereas molecules anchored to the bilayer through multiple attachment points can withstand >= 65 pN; access to this higher force regime is sufficient to probe the responses of protein-DNA interactions to force changes. As a proof-of-principle, we concurrently visualized DNA-bound fluorescently-tagged RNA polymerase while simultaneously stretching the DNA molecules. This work presents a step towards a powerful experimental platform that will enable concurrent visualization of DNA curtains while applying defined forces through optical tweezers. (C) 2012 Elsevier Inc. All rights reserved
URI
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DOI
10.1016/j.bbrc.2012.08.127
ISSN
0006-291X
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SLS_Journal Papers
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