Evidence that the herpes simplex virus type 1 uracil DNA glycosylase is required for efficient viral replication and latency in the murine nervous system.
Herpes simplex virus (HSV) encodes a uracil DNA glycosylase (UNG; UL2), which has been shown to be dispensable for normal replication of HSV-1 in cultured cells (J. Mullaney, H.W. Moss, and D.J. McGeoch, J. Gen. Virol. 70:449-454, 1989). In adult neurons, UNG activity is undetectable (F. Focher, P. Mazzarello, A. Verri, U. Hubscher, and S. Spadari, Mutat. Res. 237:65-73, 1990), suggesting that the HSV-1 UNG may play an important role in viral replication in neurons acutely and/or following reactivation. To examine the contribution of the HSV-1 UNG in vivo, two independent strain 17 Syn+ Ung- mutants, designated uB1 and uB2, were examined in a mouse model of herpetic disease. Following direct intracranial inoculation, both mutants exhibited a 10-fold reduction in neurovirulence compared with the parental strain 17 Syn+. Inoculations by a peripheral route demonstrated that the Ung- mutants were at least 100,000-fold less neuroinvasive than 17 Syn+. Replication kinetics in vivo demonstrated that uB1 and uB2 replicated less well in both the mouse peripheral and central nervous systems. Latency was established by both of the mutants in 100% of the animals examined. Following transient hyperthermia, however, the frequency of reactivation of the mutants in vivo was dramatically reduced. Restoration of the UNG locus resulted in full neurovirulence, neuroinvasiveness, and the ability to reactivate in vivo. These findings suggest that the HSV-1 UNG plays an important role during acute viral replication in vivo and possibly in the reactivation process.
Pyles
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RB
Department of Molecular Genetics, Biochemistry and Microbiology, College of Medicine, University of Cincinnati, Ohio 45267-0524.
Thompson
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RL
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D002490:Central Nervous System / Q000382:microbiology*; D003001:Cloning, Molecular; D045647:DNA Glycosylases; D006561:Herpes Simplex / Q000382:microbiology; D018259:Herpesvirus 1, Human / Q000201:enzymology* / Q000472:pathogenicity / Q000502:physiology; D008297:Male; D051379:Mice; D016296:Mutagenesis; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D017933:Peripheral Nervous System / Q000382:microbiology*; D011817:Rabbits; D051981:Uracil-DNA Glycosidase; D014774:Virulence; D014775:Virus Activation; D017735:Virus Latency; D014779:Virus Replication
D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D051981:Uracil-DNA Glycosidase