[Recombinant reverse transcriptase from Rous sarcoma virus. Kinetics and inhibition of DNA polymerase activity].
Preparations of Rous sarcoma virus reverse transcriptase isolated from a culture of E. coli HB101 (pMF14) and purified to homogeneity were used to study the steady state kinetics of DNA polymerization and inhibition of DNA-polymerase activity. DNA synthesis was examined using a system of poly(rA) as template, oligo(dT) as primer and dTTP as nucleotide substrate. Kinetic constants for steady state conditions were determined. The substrate initial velocity patterns point to an ordered mechanism which results in the formation of a ternary complex, in which the template-primer is the first to bind to the enzyme. Inhibition of the DNA-polymerase activity of the enzyme by various inhibitors was studied. Analysis of final products of the DNA-polymerase reaction revealed the presence of distribution syntheses of the DNA chain by the alpha alpha-subunit form of the enzyme.
Chernov
A P
AP
Ivanov
V A
VA
D001358:Avian Sarcoma Viruses / Q000201:enzymology*; D002384:Catalysis; D004274:DNA, Recombinant / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D019384:Nucleic Acid Synthesis Inhibitors; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D018894:Reverse Transcriptase Inhibitors
D004274:DNA, Recombinant; D019384:Nucleic Acid Synthesis Inhibitors; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase