Reverse transcriptase associated with avian sarcoma-leukosis viruses. I. Comparison of intra-virion content of multiple enzyme forms.
The RNA-dependent DNA polymerase (the reverse transcriptase) was solubilized from three related strains of avian sarcoma virus (ASV B77, ASV tsLA334, and ASV QV2) as well as avian myeloblastosis virus (AMV) and a chicken endogenous virus (RAV-O), by a combination of non-ionic detergent treatment and CsCl step-gradient centrifugation, and was subsequently separated into individual enzyme forms by poly(C)-agarose column chromatography. The newly developed two-step method allowed us to purify the three molecular forms (alpha-, alpha beta- and beta-form) of highly active enzyme rapidly and quantitatively from all the five virus strains examined. The molar ratio of the three enzyme forms differed among the virus strains: For the three sarcoma viruses, the major species was the alpha beta-form enzyme, the putative holoenzyme, and the alpha- and beta-form enzymes were less than a few percent and 15-25%, respectively, while the alpha-form enzyme content was higher for the two leukosis viruses than for the three sarcoma viruses. Both the total DNA polymerase activity and the content of the two enzyme subunits in purified virions of the three sarcoma virus was in the following order: ASV tsLA334 greater than ASV B77 greater than ASV QV2, which paralleled the virus yield at a permissive temperature in roller bottle cultures of chick embryo fibroblasts. No alteration was found in the thermolability of DNA polymerases between tsLA334, which carries ts mutations affecting both virus growth and cell-transformation, and other viruses.
D001355:Alpharetrovirus / Q000201:enzymology*; D000818:Animals; D001354:Avian Leukosis Virus / Q000201:enzymology; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002642:Chick Embryo; D002847:Chromatography, Agarose; D003370:Coturnix; D004355:Drug Stability; D005347:Fibroblasts; D006358:Hot Temperature; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D014771:Virion / Q000201:enzymology
D012194:RNA-Directed DNA Polymerase