Molecular Pharmacology, Vol 13, 496-503, Copyright © 1977 by the American Society for Pharmacology and Experimental Therapeutics

Reverse Transcription, a Probe by the Immobilized Template Poly(adenylic acid)-Agarose

¹ Department of Medical Viral Oncology, Roswell Park Memorial Institute, Buffalo, New York 14263, and School of Chemical Sciences, University of Illinois, Urbana, Illinois 62801

We have used poly(A)-agarose as an immobilized template to identify certain binding interactions which occur during polymerization of DNA and to analyze the mode of action of several drugs which inhibit oncornavirus DNA synthesis. In addition to demonstrating the binding of Moloney murine leukemia virus DNA polymerase (reverse transcriptase) to this template (both primed and unprimed), we analyzed the effects of nucleotides and several ansamycins (streptoval C, rifamycin SV, rifazone 8₂, and demethyldimethylbenzylrifampicin) on the stability of these binding interactions. The new poly(A)-agarose system allows more precise identification of the steps in reverse transcription which are targets for drugs previously known to inhibit the over-all reaction; thus this novel approach to the study of DNA polymerization on a solid matrix may afford an alternative rationale for designing antiviral drugs.

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ACKNOWLEDGMENTS We thank Dr. J. A. Huberman for his sustained interest in this work, and M. J. Eddy for technical assistance. We also thank Dr. M. Calvin and his colleagues, as well as Dr. R. C. Gallo, for their gifts of ansamycins.