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Keywords:

  • HgII ions;
  • isothermal titration calorimetry;
  • mismatched base pair;
  • nucleic acids;
  • thermodynamics

Abstract

Metal-mediated base pair formation, resulting from the interaction between metal ions and artificial bases in oligonucleotides, has been developed for its potential application in nanotechnology. We have recently found that the T:T mismatched base pair binds with HgII ions to generate a novel metal-mediated base pair in duplex DNA. The thermal stability of the duplex with the T-Hg-T base pair was comparable to that of the corresponding T:A or A:T. The novel T-Hg-T base pair involving the natural base thymine is more convenient than the metal-mediated base pairs involving artificial bases due to the lack of time-consuming synthesis. Here, we examine the specificity and thermodynamic properties of the binding between HgII ions and the T:T mismatched base pair. Only the melting temperature of the duplex with T:T and not of the perfectly matched or other mismatched base pairs was found to specifically increase in the presence of HgII ions. HgII specifically bound with the T:T mismatched base pair at a molar ratio of 1:1 with a binding constant of 106M−1, which is significantly higher than that for nonspecific metal ion–DNA interactions. Furthermore, the higher-order structure of the duplex was not significantly distorted by the HgII ion binding. Our results support the idea that the T-Hg-T base pair could eventually lead to progress in potential applications of metal-mediated base pairs in nanotechnology.