Plant Molecular Biology

, Volume 46, Issue 2, pp 161–170

Characterization of an ATP-dependent type I DNA ligase from Arabidopsis thaliana

  • You-Qiang Wu
  • Barbara Hohn
  • Alicja Ziemienowicz
Article

Abstract

Here we report the purification and biochemical characterization of recombinant Arabidopsis thaliana DNA ligase I. We show that this ligase requires ATP as a source for adenylation. The calculated Km [ATP] for ligation is 3 μM. This enzyme is able to ligate nicks in oligo(dT)/poly(dA) and oligo(rA)/poly(dT) substrates, but not in oligo(dT)/poly(rA) substrates. Double-stranded DNAs with cohesive or blunt ends are also good substrates for the ligase. These biochemical features of the purified enzyme show the characteristics typical of a type I DNA ligase. Furthermore, this DNA ligase is able to perform the reverse reaction (relaxation of supercoiled DNA) in an AMP-dependent and PPi-stimulated manner.

Arabidopsis thaliana DNA ligase intein fusion ligation relaxation 

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Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • You-Qiang Wu
    • 1
  • Barbara Hohn
    • 1
  • Alicja Ziemienowicz
    • 2
  1. 1.Friedrich Miescher-InstituteBaselSwitzerland
  2. 2.Plant Protection and Biotechnology Laboratory, Dept. of Biotechnology, Intercollegiate Faculty of BiotechnologyUniversity of GdanskGdanskPoland

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