The DNA polymerase activity of vaccinia virus 'virosomes': solubilization and properties

J Gen Virol. 1984 Apr:65 ( Pt 4):825-9. doi: 10.1099/0022-1317-65-4-825.

Abstract

Intracellular DNA-protein complexes ('virosomes') of vaccinia virus have been isolated. The solubilization of the 'virosome'-bound DNA polymerase activity was attempted by a variety of high-salt extraction procedures. The most efficient of these used 0.3 M-ammonium sulphate followed by brief sonication. The solubilized DNA polymerase activity from the 'virosomes', together with the DNA polymerases from 100000 g supernatant fluids from the cytoplasm of infected and uninfected cells were chromatographed on DEAE-cellulose and their properties compared. The 'virosome' DNA polymerase activity differed from the soluble vaccinia virus-induced DNA polymerase activity in its requirements for divalent cations and in respect of pH optimum, Km for the deoxyribonucleoside triphosphates and the effect of N-ethylmaleimide.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chlorides*
  • Chromatography, DEAE-Cellulose
  • DNA-Directed DNA Polymerase / isolation & purification
  • DNA-Directed DNA Polymerase / metabolism*
  • Deoxyribonucleoproteins / analysis*
  • Exodeoxyribonucleases / metabolism
  • HeLa Cells
  • Humans
  • Hydrogen-Ion Concentration
  • Kinetics
  • Magnesium / pharmacology
  • Magnesium Chloride
  • Manganese / pharmacology
  • Manganese Compounds*
  • Solubility
  • Vaccinia virus / analysis
  • Vaccinia virus / enzymology*
  • Viral Proteins / analysis*

Substances

  • Chlorides
  • Deoxyribonucleoproteins
  • Manganese Compounds
  • Viral Proteins
  • Magnesium Chloride
  • Manganese
  • DNA-Directed DNA Polymerase
  • Exodeoxyribonucleases
  • Magnesium
  • manganese chloride