Curved DNA molecules migrate anomalously slowly in free solution.
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 degrees 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.
Stellwagen
Earle
E
Department of Biochemistry, University of Iowa, Iowa City, IA, USA. nancy-stellwagen@uiowa.edu
Lu
Yongjun
Y
Stellwagen
Nancy C
NC
D020856:AT Rich Sequence; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics; D004247:DNA / Q000737:chemistry*; D004279:DNA, Viral / Q000737:chemistry; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D012996:Solutions
D036022:Capsid Proteins; D004279:DNA, Viral; D012996:Solutions; C062481:VP1 protein, polyomavirus; D004247:DNA