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Proc Natl Acad Sci U S A. 2013 Apr 16;110(16):E1444-51. doi: 10.1073/pnas.1218830110. Epub 2013 Apr 1.

Structural ensemble and microscopic elasticity of freely diffusing DNA by direct measurement of fluctuations.

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1
Department of Biochemistry, Stanford University, Stanford, CA 94305, USA.

Abstract

Precisely measuring the ensemble of conformers that a macromolecule populates in solution is highly challenging. Thus, it has been difficult to confirm or falsify the predictions of nanometer-scale dynamical modeling. Here, we apply an X-ray interferometry technique to probe the solution structure and fluctuations of B-form DNA on a length scale comparable to a protein-binding site. We determine an extensive set of intrahelix distance distributions between pairs of probes placed at distinct points on the surface of the DNA duplex. The distributions of measured distances reveal the nature and extent of the thermally driven mechanical deformations of the helix. We describe these deformations in terms of elastic constants, as is common for DNA and other polymers. The average solution structure and microscopic elasticity measured by X-ray interferometry are in striking agreement with values derived from DNA-protein crystal structures and measured by force spectroscopy, with one exception. The observed microscopic torsional rigidity of DNA is much lower than is measured by single-molecule twisting experiments, suggesting that torsional rigidity increases when DNA is stretched. Looking forward, molecular-level interferometry can provide a general tool for characterizing solution-phase structural ensembles.

PMID:
23576725
PMCID:
PMC3631670
DOI:
10.1073/pnas.1218830110
[Indexed for MEDLINE]
Free PMC Article
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