Department of Physics at the University of Illinois at Urbana-Champaign

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Detection of nm-scale motion caused by pico-newton forces

A DNA four-way junction (cylinders) labeled with dyes (red and green dots) are controlled by applying force via a bead trapped in a laser beam through the linker. The DNA structure interconverts between two conformations which can be detected via fluorescence resonance energy transfer (FRET) between the two dyes. How the applied force influences the interconversion frequencies can be determined by this hybrid technique combining optical trap and single molecule FRET.Researchers in Taekjip Ha's group have invented a hybrid technique to probe the dynamics of the Holliday junction, a four-stranded DNA structure that forms when damaged DNA is repaired. Understanding how DNA repairs itself is an essential step in ultimately developing effective therapies for genetic disorders.

"Based on our previous studies, we knew the Holliday junction fluctuated between two structures, but how it moved from one place to the other and what intermediates were visited along the pathway were unknown," Ha said. With this latest work, the researchers have determined that the intermediate structure is similar to a Holliday junction, bound to its own processing enzyme.

The hybrid technique combines pico-newton force control provided by an optical trap with the precise measurement capabilities of single-molecule fluorescence energy transfer. Read the paper...

The work was funded by the National Science Foundation and the National Institutes of Health. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the funding agencies.


 

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