Two avian sarcoma virus mutants with defects in the DNA polymerase-RNase H complex.
Two mutants of avian sarcoma virus which exhibit different phenotypes have been analyzed for the properties of their RNA-dependent DNA polymerase and RNase H activities. LA 338 is a complex multiple mutant with at least one lesioneach in transformation and replication functions. The purified RNA-dependent DNA polymerase-RNase H complex from the mutant is twofold more thermolabile than that from the wild-type parent. A peculiarity of this mutant is that the ability of the enzyme to respond to synthetic template-primers is lost more rapidly than is the response to native RNA as template. The mutant enzyme cannot be protected from inactivation by the addition of synthetic template-primers. LA 672 represents a different phenotype among reverse transciptase mutant, showing a "late"-acting block in replication which affects only production of progeny by infected cells grown at the nonpermissive temperature. The purified DNA polymerase-RNase H complex of LA 672 is not thermolabile; rather, progeny grown at the nonpermissive temperature yield purified enzyme with a 20-fold-reduced specific activity in both DNA polymerase and RNase H. The content of reverse transcriptase protein in such noninfectious progeny, furthermore, did not appear to be significantly diminished since immunologically active enzyme could be demonstrated in a competition test for anti-reverse transcriptase antibody and since beta and alpha subunits of reverse transcriptase could be identified after polyacrylamide gel electrophoresis of partially purified enzyme preparations. The amounts of beta and alpha from the mutant were about twofold lower.
D001358:Avian Sarcoma Viruses / Q000201:enzymology* / Q000235:genetics; D005814:Genes, Viral; D009154:Mutation; D012329:RNA, Bacterial / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*; D013696:Temperature
D012329:RNA, Bacterial; D012194:RNA-Directed DNA Polymerase; D012260:Ribonucleases