Inhibition of herpesvirus DNA synthesis by 9-beta-D-arabinofuranosyladenine in cellular and cell-free systems

Abstract

9-beta-D-Arabinofuranosyladenine 5'-triphosphate (ara-ATP) is an inhibitor both of DNA polymerase-alpha and -beta from noninfected rabbit kidney cells and of the DNA-dependent DNA polymerase induced by herpes simplex virus Type 1 (strain IES). The studies were performed with partially purified enzymes, and each of the different polymerase preparations contained only one DNA-dependent DNA polymerase species. These enzymes were inhibited in a competitive manner. The HSV-induced DNA-dependent DNA polymerase was 39-fold more sensitive to ara-ATP than was cellular DNA polymerase-beta and 116-fold more sensitive than cellular DNA polymerase-alpha. The affinity of the HSV-induced enzyme for ara-ATP was only slightly influenced by the use of different template/initiators in the enzyme assays. In intact cell systems DNA synthesis was affected by 9-beta-D-arabinofuranosyladenine (ara-A) as indicated by the reduced incorporation of deoxythymidine. In herpesvirus-(strain Lennette)-infected cells, however, ara-A shows no influence on the incorporation on deoxythymidine into cellular DNA, but it substantially reduces the incorporation into viral DNA. Ara-A itself is incorporated into both cellular and herpesviral (strain Lennette, D-316 and IES) DNA during DNA synthesis.