Mechanism of interaction of avian myeloblastosis virus reverse transcriptase with avian myeloblastosis virus RNA.
The synthesis of DNA on avian myeloblastosis virus (AMV) RNA as the primer-template using AMV reverse transcriptase in vitro has been examined as a function of the concentrations of these components, as well as a function of the ionic strenth of the assay medium. The results are consistent with the hypothesis that two types of sites exist on the AMV RNA: inactive "dead-end" sites that merely bind the enzyme, and active binding sites that lead to DNA synthesis. Velocity sedimentation studies of reverse transcriptase reveal that the enzyme becomes a dimer (or oligomer) at low salt concentrations and it is at these concentrations that the two types of sites are evident on the RNA. At high salt concentration the enzyme, which exists primarily as a monomer, is inactive with AMV RNA, although it is active when poly(rA)dT10 is used as the primer-template. We have shown that inactive sites are not due to binding of the reverse transcriptase to nicked regions or to partially denatured RNA molecules. We deduce that inactive sites are those containing incorrect 4S primer molecules. These results are discussed in terms of the mechanism of the interaction of the reverse transcriptase with AMV RNA.
D001354:Avian Leukosis Virus / Q000201:enzymology*; D009189:Avian Myeloblastosis Virus / Q000201:enzymology* / Q000378:metabolism; D001665:Binding Sites; D002474:Cell-Free System; D004279:DNA, Viral / Q000096:biosynthesis; D007700:Kinetics; D009691:Nucleic Acid Denaturation; D009994:Osmolar Concentration; D011119:Polynucleotides / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013698:Templates, Genetic
D004279:DNA, Viral; D011119:Polynucleotides; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase