The DNA polymerase activity of vaccinia virus 'virosomes': solubilization and properties.
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.
D002712:Chlorides; D002848:Chromatography, DEAE-Cellulose; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D003852:Deoxyribonucleoproteins / Q000032:analysis*; D005090:Exodeoxyribonucleases / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D015636:Magnesium Chloride; D008345:Manganese / Q000494:pharmacology; D017895:Manganese Compounds; D012995:Solubility; D014616:Vaccinia virus / Q000032:analysis / Q000201:enzymology*; D014764:Viral Proteins / Q000032:analysis*
D002712:Chlorides; D003852:Deoxyribonucleoproteins; D017895:Manganese Compounds; D014764:Viral Proteins; D015636:Magnesium Chloride; D008345:Manganese; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; D008274:Magnesium; C025340:manganese chloride