Separation of DNA-dependent DNA polymerase activities in Micrococcus radiodurans

Biochim Biophys Acta. 1977 Mar 2;475(1):23-31. doi: 10.1016/0005-2787(77)90335-5.

Abstract

DNA polymerase activities in Micrococcus radiodurans were separated into two fractions after purification more than 2000 fold. They differ in pH optimum and residual activities in the absence of a full deoxyribonucleoside triphosphates complement. NAD partly inhibited one of the activities. Both activities were eluted as a single peak on gel filtration and sedimented at the same rate on glycerol gradient centrifugation. Molecular weight 140000 was calculated from Stokes radius and sedimentation constant. Deoxyribonuclease activity was detected on one of the polymerase activities which preferentially degraded double-stranded DNA. Priming activity of nicked DNA was reduced by gamma-irradiation. These results have been related to the possible rolls in repair synthesis in vivo or DNA synthesis in permeable cells of M. radiodurans.

MeSH terms

  • DNA / radiation effects
  • DNA-Directed DNA Polymerase* / isolation & purification
  • DNA-Directed DNA Polymerase* / metabolism
  • Deoxyribonucleases / isolation & purification
  • Deoxyribonucleases / metabolism
  • Dose-Response Relationship, Radiation
  • Hydrogen-Ion Concentration
  • Kinetics
  • Micrococcus / enzymology*
  • Molecular Weight
  • NAD / pharmacology
  • Templates, Genetic

Substances

  • NAD
  • DNA
  • DNA-Directed DNA Polymerase
  • Deoxyribonucleases