DNA polymerase of murine sarcoma-leukemia virus: lack of detectable RNase H and low activity with viral RNA and natural DNA templates.
Kirsten murine sarcoma-leukemia virus (Ki-MSV[MLV]) was found to contain less RNase H per unit of viral DNA polymerase than avian Rous sarcoma virus (RSV). Upon purification by chromatography on Sephadex G-200 and subsequent glycerol gradient sedimentation the avian DNA polymerase was obtained in association with a constant amount of RNase H. By contrast, equally purified DNA polymerase of Ki-MSV(MLV) and Moloney [Mo-MSV(MLV)] lacked detectable RNase H if assayed with two homopolymer and phage fd DNA-RNA hybrids as substrates. On the basis of picomoles of nucleotides turned over, the ratio of RNase H to purified avian DNA polymerase was 1:20 and that of RNase H to purified murine DNA polymerase ranged between <1:2,800 and 5,000. Based on the same activity with poly (A).oligo(dT) the activity of the murine DNA polymerase was 6 to 60 times lower than that of the avian enzyme with denatured salmon DNA template or with avian or murine viral RNA templates assayed under various conditions (native, heat-dissociated, with or without oligo(dT) and oligo(dC) and at different template enzyme ratios). The template activities of Ki-MSV(MLV) RNA and RSV RNA were enhanced uniformly by oligo(dT) but oligo(dC) was much less efficient in enhancing the activity of MSV(MLV) RNA than that of RSV RNA. It was concluded that the purified DNA polymerase of Ki-MSV(MLV) differs from that of Rous sarcoma virus in its lack of detectable RNase H and in its low capacity to transcribe viral RNA and denatured salmon DNA. Some aspects of these results are discussed.
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000201:enzymology; D002474:Cell-Free System; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D004247:DNA; D004254:DNA Nucleotidyltransferases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D004279:DNA, Viral / Q000096:biosynthesis; D017988:Gammaretrovirus / Q000201:enzymology*; D008970:Molecular Weight; D009711:Nucleotides / Q000378:metabolism; D011119:Polynucleotides; D012367:RNA, Viral; D012260:Ribonucleases / Q000378:metabolism*; D012487:Salmonidae; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014316:Tritium
D004279:DNA, Viral; D009711:Nucleotides; D011119:Polynucleotides; D012367:RNA, Viral; D014316:Tritium; D004247:DNA; D004254:DNA Nucleotidyltransferases; D012260:Ribonucleases