Using optical tweezers to study protein-DNA interactions

Douglas E. Smith; Gregory J. Gemmen; Rachel Millin; John P. Rickgauer; Allan L. Schweitzer; Derek N. Fuller

doi:10.1117/12.618038

18 August 2005 Using optical tweezers to study protein-DNA interactions

Douglas E. Smith, Gregory J. Gemmen, Rachel Millin, John P. Rickgauer, Allan L. Schweitzer, Derek N. Fuller

Proceedings Volume 5930, Optical Trapping and Optical Micromanipulation II; 593012 (2005) https://doi.org/10.1117/12.618038
Event: Optics and Photonics 2005, 2005, San Diego, California, United States

Abstract

Mechanical manipulation of single DNA molecules can provide novel information about protein-DNA interactions. Here we review two examples studied by our group. First, we have studied the forced unraveling of nucleosomes assembled on heterogeneous DNA using core histones, the histone chaperone NAP-1, and ATP-dependent chromatin assembly and remodeling factor (ACF). We measure abrupt events releasing ~55 to 95 base pairs of DNA, which are attributable to non-equilibrium unraveling of individual nucleosomes. Wide variations observed in the unraveling force and sudden DNA re-wrapping events may have an important regulatory influence on DNA directed biochemical processes. Second, we have studied the mechanics and dynamics of single DNA looping and cleavage by "two-site" restriction enzymes. Cleavage is measured as a function of DNA tension, incubation time, and enzyme concentration, distinguishing enzymes that require DNA looping from ones that do not. Forced disruption of fixed DNA loops formed in the absence of Mg²⁺ is observed, allowing the distribution of number of loops, loop length, and disruption force to be measured as a function of time, DNA tension, and ionic conditions.

Citation Download Citation

Douglas E. Smith, Gregory J. Gemmen, Rachel Millin, John P. Rickgauer, Allan L. Schweitzer, and Derek N. Fuller "Using optical tweezers to study protein-DNA interactions", Proc. SPIE 5930, Optical Trapping and Optical Micromanipulation II, 593012 (18 August 2005); https://doi.org/10.1117/12.618038

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