Purification and characterization of the RNA-directed DNA polymerase of a primate type-D retrovirus: Mason-Pfizer virus.
The RNA-directed DNA polymerase of the primate type-D retrovirus Mason-Pfizer virus was purified using ion-exchange and affinity chromatography, and molecular sieving. The enzyme was shown to have a molecular weight of approx. 80 000 as determined by sedimentation analysis, molecular sieving and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified RNA-directed DNA polymerase retained its ability to use a heteropolymeric RNA as a template. The Mason-Pfizer virus RNA-directed DNA polymerase was also characterized as to its divalent cation preference for several synthetic primertemplates and for heteropolymeric RNA. Mg2+ was preferred as its divalent cation for all primer-templates except oligo(dG).poly(rC)m for which it prefers Mn2+. The Mason-Pfizer virus enzyme was also shown to have a pH optimum of 8-8.5 and a temperature optimum of 37-40 degrees C. The stability of the Mason-Pfizer virus RNA-directed DNA polymerase was shown to differ when measured using different primer-templates.
D000818:Animals; D009189:Avian Myeloblastosis Virus; D002413:Cations, Divalent; D002645:Chickens; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification / Q000378:metabolism*; D000882:Haplorhini; D008253:Macaca mulatta; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D009841:Oligonucleotides / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D012190:Retroviridae / Q000201:enzymology*; D013698:Templates, Genetic
D002413:Cations, Divalent; D009841:Oligonucleotides; D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium