Human cytomegalovirus. III. Virus-induced DNA polymerase.
Infection of WI-38 human fibroblasts with human cytomegalovirus (CMV) led to the stimulation of host cell DNA polymerase synthesis and induction of a novel virus-specific DNA polymerase. This cytomegalovirus-induced DNA polymerase was purified and separated from host cell enzymes by DEAE-cellulose and phosphocellulose column chromatographies. It can be distinguished from host cell enzymes by chromatographic behavior, template primer specificity, sedimentation property, and the requirement of salt for maximal activity. This virus-induced enzyme has a sedimentation coefficient of 9.2S and is found in both the nuclei and cytoplasm of virus-infected cells, but not in uninfected cells. This enzyme could efficiently use activated calf-thymus DNA, oly(dA)-oligo(dT)12-18, and poly(dC)-oligo(dG)12-18 as template primers, especially poly(dA)-oligo(dT)12-18, but it could not use poly(rA)-oligo(dT)12-18, poly(rC)-oligo(dG)12-18, or oligo(dT)12-18. The enzyme requires Mg2+ for maximal activity, is sensitive to p-hydroxymercuribenzoate, and is not a zinc metalloenzyme. In addition, the cytomegalovirus-induced DNA polymerase activity can be enhanced by adding 0.06 to 0.12 M NaCl or 0.03 to 0.06 M (NH4)2SO4 to the reaction mixture.
D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D003587:Cytomegalovirus / Q000201:enzymology*; D003593:Cytoplasm / Q000201:enzymology; D004254:DNA Nucleotidyltransferases / Q000096:biosynthesis / Q000378:metabolism; D004790:Enzyme Induction; D006801:Humans; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D008168:Lung / Q000196:embryology; D008274:Magnesium / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D011119:Polynucleotides / Q000378:metabolism; D013696:Temperature; D013698:Templates, Genetic
D006902:Hydroxymercuribenzoates; D010618:Phenanthrolines; D011119:Polynucleotides; D004254:DNA Nucleotidyltransferases; D008274:Magnesium