Comparison of virus transcription during lytic infection of the Oka parental and vaccine strains of Varicella-Zoster virus.
The attenuated Oka vaccine (V-Oka) strain of varicella-zoster virus (VZV) effectively reduces disease produced by primary infection and virus reactivation. V-Oka was developed by propagation of the Oka parental (P-Oka) strain of VZV in guinea pig and human embryo fibroblasts. Complete DNA sequencing of both viruses has revealed 63 sites that differ between P-Oka and V-Oka, 37 of which are located within 21 unique open reading frames (ORFs). Of the ORFs that differ, ORF 62 contains the greatest number (10) of mutated sites. ORF 62 encodes IE 62, the major immediate-early transactivator of virus genes, and is essential for lytic virus replication. To determine whether a disproportionate number of mutations in ORF 62 might account for virus attenuation, we compared the global pattern of V-Oka gene expression to that of P-Oka. Transcription of ORFs 62, 65, 66, and 67 was suppressed, whereas ORF 41 was elevated in V-Oka-infected cells compared to P-Oka-infected cells (P < 0.01; z test). Suppression of ORF 62, 65, and 66 transcription was confirmed by quantitative dot blot and Western blot analyses. Transient-transfection assays to determine whether mutations within V-Oka-derived IE 62 affected its ability to transactivate VZV gene promoters revealed similar IE 62 transactivation of VZV gene 20, 21, 28, 29, 65, and 66 promoters in both P-Oka and V-Oka. Together, our results indicate that mutations in V-Oka IE 62 alone are unlikely to account for vaccine virus attenuation.
Department of Neurology, University of Colorado Health Sciences Center, Denver, 80262, USA. randall.cohrs@uchsc.edu
Randall J
RJ
Cohrs
Donald H
DH
Gilden
Yasuyuki
Y
Gomi
Koichi
K
Yamanishi
Jeffrey I
JI
Cohen
D015153:Blotting, Western; D002460:Cell Line; D019433:Chickenpox Vaccine / Q000235:genetics*; D005347:Fibroblasts / Q000821:virology*; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000472:pathogenicity; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000502:physiology*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D016366:Open Reading Frames; D012333:RNA, Messenger; D012367:RNA, Viral / Q000032:analysis; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics / Q000502:physiology*; D014764:Viral Proteins / Q000032:analysis; D014779:Virus Replication / Q000235:genetics*
D019433:Chickenpox Vaccine; C074221:IE62 protein, Human herpesvirus 3; D017874:Immediate-Early Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D014764:Viral Proteins