Environmental influences on DNA superhelicity. The effect of ionic strength on superhelix conformation in solution.
The techniques of small-angle X-ray scattering and analysis that have been developed by the authors are used to investigate the influence of ionic strength on the superhelical conformation of native COP608 plasmid DNA in solution. For salt concentrations below 0.1 M, the superhelicity is partitioned between twisting (Tw) and writhing (Wr) in the ratio delta Tw/Wr = 2. Near the physiological salt concentration, [Na+] = 0.2 M, a co-operative transition is observed in which the pitch angle of the toroidal superhelix is drastically decreased. This results in an almost complete relaxation of writhe. At salt concentrations in excess of the threshold for this transition, the superhelical partitioning occurs in the ratio delta Tw/Wr greater than 25. Energetic considerations support the suggestion that this transition results from co-operative, superhelical B to Z transconformation reactions at susceptible sites. A method is discussed that will enable the direct measurement of this secondary structural transition by means of X-ray scattering.
Brady
G W
GW
Center for Laboratories & Research, New York State Department of Health, Albany 12201.
Satkowski
M
M
Foos
D
D
Benham
C J
CJ
D004278:DNA, Superhelical; D009690:Nucleic Acid Conformation; D009994:Osmolar Concentration; D012542:Scattering, Radiation; D014965:X-Rays
D004278:DNA, Superhelical