Exchange of counterions in DNA condensation.
We measured the fluorescence intensity of DNA-bound fluorescent dyes YO-PRO-1 (oxazole yellow) and YOYO-1 (dimer of oxazole yellow) at various spermidine concentrations to determine how counterions on DNA are exchanged in the process of DNA condensation. A decrease of fluorescence intensity was observed with an increase of spermidine. Considering the chemical equilibrium under the competition between the dye and spermidine for counterion condensation on DNA, the theoretical curve well describes the decrease of the fluorescence intensity. These results indicate that dyes are exchanged for spermidine at the binding site on DNA; that is, the exchange of counterions occurs. The parameters associated with the decrease of the fluorescence intensity show that the relative affinity of the dye and spermidine for DNA depends on the state of DNA. Moreover, YOYO-1 prevents the DNA condensation, but the effect of YO-PRO-1 on the condensation is very slight, though both dyes intercalate for DNA; the high affinity of YOYO-1 compared to YO-PRO-1 enables prevention of the condensation.
Murayama
Yoshihiro
Y
Department of Physics, University of Tokyo, Tokyo 113-0033, Japan. murayama@daisy.phys.s.u-tokyo.ac.jp
Sano
Masaki
M
D010582:Bacteriophage lambda; D001665:Binding Sites; D004279:DNA, Viral / Q000737:chemistry*; D005456:Fluorescent Dyes; D013050:Spectrometry, Fluorescence; D013095:Spermidine / Q000494:pharmacology
D004279:DNA, Viral; D005456:Fluorescent Dyes; D013095:Spermidine