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Proceedings Paper

Using optical tweezers to study protein-DNA interactions
Author(s): Douglas E. Smith; Gregory J. Gemmen; Rachel Millin; John P. Rickgauer; Allan L. Schweitzer; Derek N. Fuller
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Paper 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.

Paper Details

Date Published: 19 August 2005
PDF: 10 pages
Proc. SPIE 5930, Optical Trapping and Optical Micromanipulation II, 593012 (19 August 2005); doi: 10.1117/12.618038
Show Author Affiliations
Douglas E. Smith, Univ. of California, San Diego (United States)
Gregory J. Gemmen, Univ. of California, San Diego (United States)
Rachel Millin, Univ. of California, San Diego (United States)
John P. Rickgauer, Univ. of California, San Diego (United States)
Allan L. Schweitzer, Univ. of California, San Diego (United States)
Derek N. Fuller, Univ. of California, San Diego (United States)


Published in SPIE Proceedings Vol. 5930:
Optical Trapping and Optical Micromanipulation II
Kishan Dholakia; Gabriel C. Spalding, Editor(s)

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