Complexing reverse transcriptase with polyspermine-ribonuclease.
Polyspermine-ribonuclease (Mr approximately 17 000) and the enzyme transcriptase from Rauscher-leukaemia virus (Mr approximately 70 000) form a complex Mr approx. 160 000) such that the molar ratio of polyspermine-ribonuclease to reverse transcriptase is 5:1. The most favourable condition for complex-formation is in a solution consisting of 0.01 M-Tris/HCl buffer, pH 7.5, 0.25 M-KCl and 1 mM-Mn2+ at 37 degrees C. The association of the two enzymes retains full RNAase activity, but reverse-transcriptase activity is completely inhibited when ribonuclease-sensitive polymers such as (dG)12 x (rC)n or viral 70S RNA are used as primer templates.
D008970:Molecular Weight; D009097:Multienzyme Complexes; D012194:RNA-Directed DNA Polymerase; D012260:Ribonucleases; D013096:Spermine / Q000031:analogs & derivatives; D014758:Viral Core Proteins; D014764:Viral Proteins
D009097:Multienzyme Complexes; D014758:Viral Core Proteins; D014764:Viral Proteins; D013096:Spermine; D012194:RNA-Directed DNA Polymerase; D012260:Ribonucleases