PROPERTIES OF HERPES-SIMPLEX VIRUS-DNA POLYMERASE AND CHARACTERIZATION OF ITS ASSOCIATED EXONUCLEASE ACTIVITY

Herpes simplex virus (HSV) DNA polymerase was isolated on a large‐scale from African green monkey kidney cells infected with HSV type 1 (HSV‐1) strain Angelotti. After DNA‐cellulose chromatography the enzyme showed a specific activity of 48000 units/mg protein. Three major single polypeptides with molecular weights of 144000, 74000 and 29000 were copurified with the enzyme activity at the DNA‐cellulose step. By its chromatographic behavior and by template studies, the HSV DNA polymerase activity was clearly distinguishable from cellular α, β and γ DNA polymerase activities. Two exonucleolytic activities were found in the DNA‐cellulose enzyme preparation. The main exonucleolytic activity, which degraded both single‐stranded and double‐stranded DNA to deoxynucleoside 5′‐monophosphates, was separated by subsequent velocity sedimentation. The remaining exonucleolytic activity was not separable from the HSV DNA polymerase by several chromatographic steps and by velocity sedimentation at high ionic strength. This novel exonuclease and HSV DNA polymerase were equally sensitive both to phosphonoacetic acid and Zn2+ ions, inhibitors of the viral polymerase. Similar to the 3′‐to‐5′‐exonuclease of procaryotic DNA polymerases and mammalian DNA polymerase δ, the HSV‐polymerase‐associated exonuclease catalyzed the removal of 3′‐terminal nucleotides from the primer/template as well as the template‐dependent conversion of deoxynucleoside triphosphates to monophosphates. Copyright © 1979, Wiley Blackwell. All rights reserved