AI Article Synopsis
- A transient assay system was utilized to identify VZV genes that can activate early (tk promoter) and late (gpI promoter) gene expression.
- In Vero cells, cotransfection with specific VZV DNA fragments and a CAT reporter construct showed significant increases in CAT expression, particularly with VZV genes ORF4 and ORF62.
- The study also revealed a VZV-encoded repressor involving ORF60 and ORF61 that inhibits p1tkCAT expression and can modulate the activation effects of ORF4 and ORF62.
Article Abstract
A transient assay system was used to identify varicella-zoster virus (VZV)-encoded genes whose products are able to activate the expression of an early gene promoter, the thymidine kinase (tk) promoter, and a late gene promoter, and the glycoprotein I (gpI) promoter. Vero cells were cotransfected with individual cloned DNA fragments spanning the entire VZV genome and with the recombinant construct p1tkCAT which contained the chloramphenicol acetyl transferase (CAT) gene under the control of putative regulatory sequences. Five- to 20-fold increases in the expression p1tkCAT was observed in cotransfections with plasmids containing VZV open reading frame (ORF)4 (map location 0.02-0.03) or ORF62 (0.82-0.86). Expression of p68CAT (contains -682 to +222 bp relative to the AUG of gpI) was also enhanced by the products of ORF4 and ORF62. Synergy between ORF4 and ORF62 products was observed in the activation of p68CAT, resulting in a 22-fold increase in CAT activity. RNA analysis indicated that activation of these promoters was at the transcriptional level. A VZV-encoded "repressor" sequence, containing ORF60 and ORF61, was also identified which repressed expression of p1tkCAT and modulated its activation by ORF4 and ORF62.
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