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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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On-off Regulation of 3' Exonuclease Excision to DNA Polymerization by Exo+ Polymerase

  • Zhang, Jia (Genomapping Inc.) ;
  • Li, Kai (Institute of Life Science and Biotechnology, Jinan University)
  • Received : 2003.02.07
  • Accepted : 2003.03.04
  • Published : 2003.11.30
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Abstract

The role of 3' exonuclease excision in DNA polymerization was evaluated in primer extensions using 3' allele-specific primers that had exonuclease-digestible and exonuclease-resistant 3' termini. With exonuclease-digestible unmodified 3' mismatched primers, the exo+ polymerase yielded template-dependent products. Using exonuclease-resistant 3' mismatched primers, no primer-extended product resulted from exo+ polymerase. As a control, polymerase without proofreading activity yielded primer-dependent products from 3' mismatched primers. These data indicated that a successful removal of the mismatch is required for DNA polymerization from the 3' mismatched primers by exo+ polymerase. In addition to the well-known proofreading from this mismatch removal, the premature termination in DNA polymerization, due to the failure of the efficient removal of the mismatched nucleotides, worked as an off-switch in maintaining the high fidelity in DNA replication from exo+ polymerase.

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