Characteristics of migration patterns of DNA oligomers in gels and the relationship to the question of intrinsic DNA bending.
We have developed a methodology that is capable of quantitatively describing the electrophoretic mobility patterns of oligomeric B-DNA through polyacrylamide gels (PAG) in the presence of varying concentration of the organic solvent 2-methyl-2,4-pentanediol (MPD), used routinely to induce DNA crystallization. The model includes the ion atmosphere and its polarization, electrostatic excluded volume, hydrodynamic interactions, and fluctuation effects that characterize the overall size of the migrating polyion. Using this model, and by critically examining the mobility patterns of linear random-sequence B-DNA molecules in PAG as a function of MPD, we address the question of the discrepancy between current models used to explain the molecular origins of A-tract-induced DNA bending. Direct analysis of the mobility of B-DNA oligomers on PAG, and comparison to the mobility of A-tract-containing oligomers, shows a significant apparent effect of MPD on the mobility of generic B-DNA sequences, which is larger than the effect on A-tract-containing oligomers. The effect is chain-length dependent, especially at lower MPD concentration. Thus, the apparent reduction in gel mobility, as a function of MPD, is not unique to A-tract regions or A-tract-containing molecules. However, our analysis suggests that MPD molecules are probably excluded from the surface of both B-DNA and A-tract molecules. This is supported by circular dichroism studies on A-tract and B-DNA molecules in solutions containing various MPD concentrations.
Department of Biology, Technion-Israel Institute of Technology, Technion City, Haifa 32000, Israel. bitali@tx.technion.ac.il
Tali E
TE
Haran
Ilana
I
Cohen
Alexander
A
Spasic
Kevin
K
Yang
Udayan
U
Mohanty
D000180:Acrylic Resins / Q000737:chemistry*; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006018:Glycols / Q000737:chemistry; D008956:Models, Chemical; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry
D000180:Acrylic Resins; D006018:Glycols; D009841:Oligonucleotides; C016680:polyacrylamide gels; D004247:DNA; C011480:hexylene glycol