Mode of inhibition of herpes simplex virus DNA polymerase by phosphonoacetate.
Phosphonoacetate is a highly specific inhibitor of herpes simplex virus-induced DNA polymerase. Sensitivity of herpesvirus type 1 or type 2 induced DNA polymerase to the drug was similar. However, DNA polymerases from other sources such as the host cells (Wi-38), Micrococcus luteus, and hepatitis B virus were highly resistant. In addition, Escherichia coli RNA polymerase and reverse transcriptase of Rous sarcoma virus were also insensitive to the drug. Enzyme kinetic studies showed that inhibition was noncompetitive with respect to deoxyribonucleotide triphosphates. The Ki value was about 0.45 muM. The apparent Km values for dTTP, dATP, dCTP, and dGTP were 0.71, 0.75, 0.42, and 0.39 muM, respectively. The base composition of template has no profound effect on the extent of inhibition. The drug caused uncompetititve inhibition with respect to template which indicated that phosphonoacetate did not bind directly to template DNA. Results are presented which suggest that phosphonoacetate did not affect the formation of the enzyme-DNA complex but probably inhibited the elongation step of DNA polymerase reaction.
D000085:Acetates / Q000494:pharmacology*; D001358:Avian Sarcoma Viruses / Q000201:enzymology; D004254:DNA Nucleotidyltransferases / Q000037:antagonists & inhibitors*; D004279:DNA, Viral / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors; D003854:Deoxyribonucleotides / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology; D006515:Hepatitis B virus / Q000201:enzymology; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008837:Micrococcus / Q000201:enzymology; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D011188:Potassium / Q000494:pharmacology; D011485:Protein Binding; D018894:Reverse Transcriptase Inhibitors; D018139:Simplexvirus / Q000201:enzymology*; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000494:pharmacology
D000085:Acetates; D004279:DNA, Viral; D003854:Deoxyribonucleotides; D009943:Organophosphorus Compounds; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; D004254:DNA Nucleotidyltransferases; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium; D011188:Potassium