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.