| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1994: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1993: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Research Abstract |
1. The protein kinase encoded by the US3 gene of herpes simplex virus type 2 (HSV-2) was purified more than 1000-fold from the post-ribosomal supernatant of infected Vero cells, and the final preparation contained one major protein of apparent molecular weight 66K,which was phosphorylated in the autophosphorylation reaction. The HSV-2 protein kinase was relatively resistant to high concentrations of salt, and when the substrate specificity was investigated using synthetic oligopeptides, the peptides containing arginyl residues on the amino-terminal side of the target residue were found to be the best substrates for the US3 protein kinase.
2. When permeabilized cells were labelled with [gamma-^<32>P] ATP under the optimum conditions for the US3 protein kinase, the most striking difference between wild-type and US3-inactivated virus-infected cells was observed in the phosphorylation of proteins ranging in Mr values from 14K to 21K.The results indicate that a tegment phosphoprotein encoded by the US9 gene may be a target of the enzyme.Similar studies demonstrate that the alkaline nuclease encoded by UL12 was also a major target of the US3 protein kinase, and suggest that phosphorylation by the protein kinase may be involved in the stability/activity of the alkaline nuclease in murine macrophages.
3. We have studied the pathogenic and latency/reactivation potential of a herpes simplex virus type 1 (HSV-1) variant (N38) which has a deletion of four genes, US9,10,11 and 12, in the short unique region of the HSV-1 genome. N38 was a pathogenic as the parental wild-type virus following intracerebral infection of mice and replicated with wild-type kinetics in the brain. There was no significant difference between two viruses in the frequency of reactivation or in reactivation time. The results indicate that these four genes are dispensable for its neurovirulence and latency in mice.
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