The intrinsic curvature of seven 98 bp DNA molecules containing up to four centrally located A6-tracts has been measured by gel and capillary electrophoresis as a function of the number and arrangement of the A-tracts. At low cation concentrations, the electrophoretic mobility observed in polyacrylamide gels and in free solution decreases progressively with the increasing number of phased A-tracts, as expected for DNA molecules with increasingly curved backbone structures. Anomalously slow electrophoretic mobilities are also observed for DNA molecules containing two pairs of phased A-tracts that are out of phase with each other, suggesting that out-of-phase distortions of the helix backbone do not cancel each other out. The mobility decreases observed for the A-tract samples are due to curvature, not cation binding in the A-tract minor groove, because identical free solution mobilities are observed for a molecule with four out-of-phase A-tracts and one with no A-tracts. Surprisingly, the curvature of DNA A-tracts is gradually lost when the monovalent cation concentration is increased to ∼200 mM, regardless of whether the cation is a hydrophilic ion like Na+, NH4+, or Tris+ or a hydrophobic ion like tetrabutylammonium. The decrease in A-tract curvature with increasing ionic strength, along with the known decrease in A-tract curvature with increasing temperature, suggests that DNA A-tracts are not significantly curved under physiological conditions.
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http://dx.doi.org/10.1021/bi400118m | DOI Listing |
Biophys J
January 2025
Department of Mathematics and Statistics, Haverford College, Haverford, Pennsylvania. Electronic address:
We apply the Laplace approximation to a mathematical formulation of DNA cyclization J factors, leading to a formula that involves energies of local minima of the DNA energy, factors coming from the Hessian of the energy near each minimum, and geometric factors arising from the orientational portion of J. The approximation is derived in a quite general setting that encompasses both rigid base and rigid basepair models common in the literature. The approximation is applied to several families of 200-400 bp DNA, some relatively straight (fragments of λ-phage) and others quite bent (constructs that include up to 10 A tracts).
View Article and Find Full Text PDFNanoscale
October 2024
School of Physics, Engineering and Technology, University of York, York, YO10 5DD, UK.
Periodic sequences in phase with DNA helical shape are prevalent in genomes due to their capacity to modulate DNA elasticity on a global scale. However, how this occurs is not well understood. We use all-atom molecular dynamics simulations on 40 bp DNA fragments to assess the effect of periodicity on bending, twisting, and stretch elasticity.
View Article and Find Full Text PDFElectrophoresis
March 2024
Department of Biochemistry, University of Iowa, Iowa City, Iowa, USA.
Capillary electrophoresis has been used to measure the free solution mobilities of a series of 26-base pair (bp) DNA oligomers containing two phased A4T1in-tracts embedded in flanking sequences containing 0 to 11 additional AT bps. A random-sequence 26-bp oligomer with 12 isolated AT bps was used as the reference. Mobility ratios (A-tract/reference) were measured in background electrolytes (BGEs) containing mixtures of small monovalent cations and tetrabutylammonium (TBA ) or tetrapropylammonium (TPA ) ions.
View Article and Find Full Text PDFElectrophoresis
September 2023
Department of Biochemistry, University of Iowa, Iowa City, Iowa, USA.
The free solution mobilities of 26-base pair (bp) DNA oligomers containing A-tracts with and without internal ApT steps have been measured by capillary electrophoresis, using the mobility of a 26-bp random-sequence oligomer as a reference. The background electrolytes (BGEs) contained mixtures of Li and tetrapropylammonium (TPA ) ions, keeping the total cation concentration constant at 0.3 M.
View Article and Find Full Text PDFNucleic Acids Res
December 2022
Institut f. Synthetische Mikrobiologie, Heinrich-Heine Universität Düsseldorf, Universitätsstrasse 1, 40225 Düsseldorf, Germany.
In cyanobacteria DNA supercoiling varies over the diurnal cycle and is integrated with temporal programs of transcription and replication. We manipulated DNA supercoiling in Synechocystis sp. PCC 6803 by CRISPRi-based knockdown of gyrase subunits and overexpression of topoisomerase I (TopoI).
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