Purification and characterization of varicella-zoster virus-induced DNA polymerase.
Infection of WI-38 human fibroblasts with varicella-zoster virus led to the stimulation of host cell DNA polymerase synthesis and induction of a new virus-specific DNA polymerase. This virus-induced DNA polymerase was partially purified and separated from host cell enzymes by DEAE-cellulose and phosphocellulose column chromatographies. This virus-induced enzyme could be distinguished from host cell enzyme by its chromatographic behavior, template specificity, and its requirement of salt for maximal activity. The enzyme could efficiently use poly(dC).oligo(dG)12-18 as well as poly(dA).oligo(dT)12-18 as template-primers. It required Mg2+ for maximal polymerization activity and was sensitive to phosphonoacetic acid, to which host alpha- and beta-DNA polymerase were relatively resistant. In addition, this induced DNA polymerase activity was enhanced by adding 60 mM (NH4)2SO4 to the reaction mixture.
D000645:Ammonium Sulfate / Q000494:pharmacology; D002460:Cell Line; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D004790:Enzyme Induction; D014645:Herpesvirus 3, Human / Q000201:enzymology*; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D010746:Phosphonoacetic Acid / Q000494:pharmacology; D011089:Polydeoxyribonucleotides / Q000378:metabolism
D006902:Hydroxymercuribenzoates; D010618:Phenanthrolines; D011089:Polydeoxyribonucleotides; D004259:DNA-Directed DNA Polymerase; D008274:Magnesium; D010746:Phosphonoacetic Acid; D000645:Ammonium Sulfate