Inhibition of DNA polymerases alpha and gamma by rifamycin derivative AF/013.
The nature of the inhibitory effects of rifamycin derivative AF/013 (O-n-octyloxime of rifamycin SV) on DNA polymerases alpha and gamma were studied. Lineweaver-Burk analysis of the inhibition of DNA polymerases with respect to a substrate and template-primer showed a different mode of inhibition by AF/013 for each: the inhibition of DNA polymerase gamma was competitive with both dTTP and poly(rA)-oligo(dT), while that of DNA polymerase alpha was competitive with activated calf thymus DNA and non-competitive with dTTP. Further analysis of the competitive mode of the inhibition of DNA polymerase alpha, using poly(dT)-oligo(rA) as a template-primer, demonstrated that the primer molecule competed with AF/013. A change of effective divalent metal ion (Mn2+ in place of Mg2+) in the reaction mixture did not alter this competitive mode of inhibition with respect to the template-primer. The results of experiments to obtain further insight into the mechanism of drug-enzyme interaction suggest that AF/013 binds tightly to DNA polymerase alpha, and inhibits the process of chain elongation with DNA polymerases alpha and gamma.
D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D001667:Binding, Competitive; D002286:Carcinoma, Ehrlich Tumor; D002460:Cell Line; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors*; D004258:DNA Polymerase III / Q000037:antagonists & inhibitors*; D007700:Kinetics; D019384:Nucleic Acid Synthesis Inhibitors; D011485:Protein Binding; D012294:Rifamycins / Q000494:pharmacology*
D019384:Nucleic Acid Synthesis Inhibitors; D012294:Rifamycins; C006019:AF 013; D004257:DNA Polymerase II; D004258:DNA Polymerase III