Phenotypic properties of herpes simplex virus 1 containing a derepressed open reading frame P gene.
Open reading frame P (ORF P) maps in the viral DNA sequences transcribed during latency and is located antisense to the gamma 1 34.5 gene. Earlier studies have shown that the expression of ORF P is repressed by an infected cell protein no. 4 binding site straddling the transcription initiation site. We have made monospecific polyclonal antibodies to the protein and constructed a virus, designated ORF P++, in which the infected cell protein no. 4 binding site has been mutagenized, thereby allowing full expression of an unmodified ORF P gene from its natural promoter. We report the following findings. (i) The native protein forms multiple bands on denaturing polyacrylamide gels suggestive of extensive processing and aggregation of the protein; (ii) the protein accumulates in the nucleus in rod-shaped structures perpendicular to the axis of attachment of the infected cell to the solid matrix; (iii) the virus was highly attenuated on inoculation into mice by the intracerebral or ocular route, and virus was not recovered upon explantation of trigeminal ganglia; (iv) although protein synthesis was not prematurely shut off in the human neuroblastoma cell line SK-N-SH, gamma 1 34.5 protein was not detected in immunoblasts. Analyses of electrophoretically separated denatured RNAs indicated that in cells infected with the ORF P++ virus, there was a large increase in the amount of ORF P RNA and a corresponding decrease in the amount of gamma 1 34.5 RNA. We conclude that either the overproduction of ORF P protein blocks the expression of some herpes simplex virus 1 genes or derepression of the transcription of ORF P has a negative effect on the transcription of the antisense gamma 1 34.5 RNA.
Lagunoff
M
M
Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, Illinois 60637, USA.
Randall
G
G
Roizman
B
B
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001665:Binding Sites / Q000235:genetics; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015967:Gene Expression Regulation, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism; D051379:Mice; D008808:Mice, Inbred CBA; D016296:Mutagenesis; D016366:Open Reading Frames; D010641:Phenotype; D011817:Rabbits; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D014709:Vero Cells; D014764:Viral Proteins / Q000032:analysis / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D014779:Virus Replication
D000914:Antibodies, Viral; D017874:Immediate-Early Proteins; D014764:Viral Proteins; C066469:gamma 34.5 protein, Human herpesvirus 1; C048925:herpes simplex virus, type 1 protein ICP4