Mechanism of degradation of duplex DNA by the DNase induced by herpes simplex virus.
Reaction intermediates formed during the degradation of linear PM2, T5, and lambda DNA by herpes simplex virus (HSV) DNase have been examined by agarose gel electrophoresis. Digestion of T5 DNA by HSV type 2 (HSV-2) DNase in the presence of Mn(2+) (endonuclease only) gave rise to 6 major and 12 minor fragments. Some of the fragments produced correspond to those observed after cleavage of T5 DNA by the single-strand-specific S1 nuclease, indicating that the HSV DNase rapidly cleaves opposite a nick or gap in a duplex DNA molecule. In contrast, HSV DNase did not produce distinct fragments upon digestion of linear PM2 or lambda DNA, which do not contain nicks. In the presence of Mg(2+), when both endonuclease and exonuclease activities of the HSV DNase occur, most of the same distinct fragments from digestion of T5 DNA were observed. However, these fragments were then further degraded preferentially from the ends, presumably by the action of the exonuclease activity. Unit-length lambda DNA, EcoRI restriction fragments of lambda DNA, and linear PM2 DNA were also degraded from the ends by HSV DNase in the same manner. Previous studies have suggested that the HSV exonuclease degrades in the 3' --> 5' direction. If this is correct, and since only 5'-monophosphate nucleosides are produced, then HSV DNase should "activate" DNA for DNA polymerase. However, unlike pancreatic DNase I, neither HSV-1 nor HSV-2 DNase, in the presence of Mg(2+) or Mn(2+), activated calf thymus DNA for HSV DNA polymerase. This suggests that HSV DNase degrades both strands of a linear double-stranded DNA molecule from the same end at about the same rate. That is, HSV DNase is apparently capable of degrading DNA strands in the 3' --> 5' direction as well as in the 5' --> 3' direction, yielding progressively smaller double-stranded molecules with flush ends. Except with minor differences, HSV-1 and HSV-2 DNases act in a similar manner.
D010582:Bacteriophage lambda; D001435:Bacteriophages; D004279:DNA, Viral / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D018139:Simplexvirus / Q000201:enzymology*; D013698:Templates, Genetic
D004279:DNA, Viral; D008345:Manganese; D004259:DNA-Directed DNA Polymerase; D003851:Deoxyribonucleases; D008274:Magnesium