Fluorescence-Force Spectroscopy Maps Two-Dimensional Reaction Landscape of the Holliday Junction

Despite the recent advances in single-molecule manipulation techniques, purely mechanical approaches cannot detect subtle conformational changes in the biologically important regime of weak forces. We developed a hybrid scheme combining force and fluorescence that allowed us to examine the effect of...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2007-10, Vol.318 (5848), p.279-283
Main Authors: Hohng, Sungchul, Zhou, Ruobo, Nahas, Michelle K, Yu, Jin, Schulten, Klaus, Lilley, David M.J, Ha, Taekjip
Format: Article
Language:English
Subjects:
Bacteriophage lambda
Biological and medical sciences
Biophysical Phenomena
Biophysics
Biosensors
Conformity
Coordinate systems
Cruciform DNA
DNA
DNA, Cruciform - chemistry
DNA, Viral - chemistry
Enzymes
Fluorescence
Fluorescence Resonance Energy Transfer
Fundamental and applied biological sciences. Psychology
Interactions. Associations
Intermolecular phenomena
Mechanics
Molecular biology
Molecular biophysics
Molecules
Nucleic Acid Conformation
Nucleic acids
Optical Tweezers
Soil chemistry
Spectrum analysis
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Summary:Despite the recent advances in single-molecule manipulation techniques, purely mechanical approaches cannot detect subtle conformational changes in the biologically important regime of weak forces. We developed a hybrid scheme combining force and fluorescence that allowed us to examine the effect of subpiconewton forces on the nanometer scale motion of the Holliday junction (HJ) at 100-hertz bandwidth. The HJ is an exquisitely sensitive force sensor whose force response is amplified with an increase in its arm lengths, demonstrating a lever-arm effect at the nanometer-length scale. Mechanical interrogation of the HJ in three different directions helped elucidate the structures of the transient species populated during its conformational changes. This method of mapping two-dimensional reaction landscapes at low forces is readily applicable to other nucleic acid systems and their interactions with proteins and enzymes.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.1146113