Replication, latent infection, and reactivation in neuronal culture with a herpes simplex virus thymidine kinase-negative mutant.
Herpes simplex virus type 1 (HSV-1) mutant viruses lacking functional viral thymidine kinase activity are reported to be incapable of replication in neurons. To investigate the role of viral thymidine kinase (TK) activity in the HSV-1 infection of the neuron, we studied a thymidine kinase-negative (TK-) mutant virus engineered to eliminate TK function without affecting the other known transcripts encoded in this region of the genome. Studies using the mouse eye model demonstrated that the mutant behaved as is reported for other TK- viruses: DNA of the mutant virus was detected in the ganglia during the latent infection by polymerase chain reaction, but virus did not reactivate after explantation of the ganglia. Utilizing the neuronal cultures, we investigated the ability of the mutant virus to replicate in neurons and the capacity of the mutant virus to establish latency and reactivate. With a low multiplicity of infection (m.o.i.), replication of the TK- mutant virus in sensory neurons in culture was significantly delayed compared to that of the wild-type virus. However, when a high m.o.i. was used, the mutant and the wild-type viruses replicated with similar kinetics. The TK- mutant virus was capable of establishment of latency and reactivation from the latent infection in sensory neurons in culture. These data suggest that HSV-1 thymidine kinase activity facilitates viral replication, but that TK activity is not essential for either replication or reactivation from latent infections in neurons in vitro.
Wilcox
C L
CL
Department of Microbiology, University of Colorado School of Medicine, Denver 80262.
Crnic
L S
LS
Pizer
L I
LI
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D003315:Cornea / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009474:Neurons / Q000382:microbiology*; D018139:Simplexvirus / Q000235:genetics / Q000502:physiology*; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism; D012668:Trigeminal Ganglion / Q000382:microbiology; D010948:Viral Plaque Assay; D014775:Virus Activation; D014779:Virus Replication
D013937:Thymidine Kinase