Purification and immunological characterization of reverse transcriptase associated with hepatitis non-A, non-B

Abstract

The infectious viral particles present in a hepatitis non-A, non-B patient serum (inoculum I) were sedimented by centrifugation. Following detergent disruption, the particle-associated reverse transcriptase in the sediment was fractionated by affinity chromatography and characterized by immunoblot analyses and radioimmunoprecipitation. By using specific antibodies to simian sarcoma virus reverse transcriptase, a cross-reactive protein of 80,000 daltons was detected in inoculum I, but not in a control serum. During affinity chromatography of the viral lysate on oligo(dC) cellulose (either stepwise or gradient elution), the reverse transcriptase was eluted by 0.2–0.3 m KC1. The reverse transcriptase thus purified was immunoprecipitated by antisera to reverse transcriptase from type C mammalian retroviruses, including simian sarcoma virus (SSV), RD114, baboon endogenous virus (BaEV), and Rauscher leukemia virus (RLV). No significant immunoprecipitate was obtained with antisera to reverse transcriptase from avian myeloblastosis virus (AMV) or type B and type D viruses. These results indicate that the reverse transcriptase purified from hepatitis non-A, non-B serum shares one or more determinants with other type C mammalian virus reverse transcriptases.