Published online 5 August 2005
Article |
Curved DNA molecules migrate anomalously slowly in free solution
Department of Biochemistry, University of Iowa Iowa City, IA, USA 1Department of Internal Medicine, University of Iowa Iowa City, IA, USA
*To whom correspondence should be addressed. Tel: +1 319 335 7896; Fax: +1 319 335 9570; Email: nancy-stellwagen{at}uiowa.edu
Received May 2, 2005. Revised July 15, 2005. Accepted July 15, 2005.
The electrophoretic mobility of a curved DNA restriction fragment taken from the VP1 gene in the SV40 minichromosome has been measured in polyacrylamide gels and free solution, using capillary electrophoresis. The 199 bp restriction fragment has an apparent bend angle of 46 ± 2° located at SV40 sequence position 1922 ± 2 bp [Lu Y.J., Weers B.D. and Stellwagen N. C. (2005) Biophys. J., 88, 1191–1206]. The ‘curvature module’ surrounding the apparent bend center contains five unevenly spaced A- and T-tracts, which are responsible for the observed curvature. The parent 199 bp fragment and sequence mutants containing at least one A-tract in the curvature module migrate anomalously slowly in free solution, as well as in polyacrylamide gels. Hence, the anomalously slow mobilities observed for curved DNA molecules in polyacrylamide gels are due in part to their anomalously slow mobilities in free solution. Analysis of the gel and free solution mobility decrements indicates that each A- or T-tract contributes independently, but not equally, to the curvature of the 199 bp fragment and its A-tract mutants. The relative contribution of each A- or T-tract to the observed curvature depends on its spacing with respect to the first A-tract in the curvature module.