Radioimmunoassay for mammalian type C viral reverse transcriptase

Abstract

Radioimmunological techniques were applied to the analysis of reverse transcriptase of mammalian type C RNA viruses. The polymerase of Rauscher mouse leukemia virus was purified by ion exchange and sequential affinity chromatography. Radioimmunoassays that utilized the viral enzyme as a probe detected as little as 1 ng of purified polymerase. No cross-reactivity could be demonstrated between the reverse transcriptase and other known virus-coded proteins. By comparing the immunological reactivity of the purified enzyme with the reactivity of detergent-disrupted virions, Rauscher mouse leukemia virus was shown to contain the antigenic equivalent of 40 molecules of reverse transcriptase. In a homologous competition immunoassay, the Rauscher viral enzyme demonstrated type-specific antigenic determinants, which distinguish it from other mouse type C viral polymerases. In a broadly reactive interspecies immunoassay, the reverse transcriptases of a number of mammalian type C viruses were cross-reactive, indicating their shared antigenic determinants. Various treatments that inhibit or inactivated DNA polymerase activity had little or no effect on the immunological properties of the enzyme. Thus, radioimmunoassays should be useful in the search for type C viral reverse transcriptase as a marker of subviral expression.