Activities of herpes simplex virus type 1 (HSV-1) ICP4 genes specifying nonsense peptides

Abstract

Synthetic oligonucleotide linkers containing translational termination codons in all possible reading frames were inserted at various positions in the cloned gene encoding the herpes simplex virus type 1 (HSV-1) immediate-early regulatory protein, ICP4. It was determined that the amino-terminal 60 percent of the ICP4 gene was sufficient for trans-induction of a thymidine kinase promoter-CAT chimera (pTKCAT) and negative regulation of an ICP4 promoter-CAT chimera (pIE3CAT); however, it was relatively inefficient in complementing an ICP4 deletion mutant. The amino-terminal ninety amino acids do not appear to be required for infectivity as reflected by the replication competence of a mutant virus containing a linker insertion at amino acid 12. The size of the ICP4 molecule expressed from the mutant virus was consistent with translational restart at the next methionine codon corresponding to amino acid 90 of the deduced ICP4 amino acid sequence.

Hepatoma-specific antitumor activity of an albumin enhancer/promoter regulated herpes simplex virus in vivo

“…Hep3B, HeLa and Vero cells were obtained from American Type Culture Collection (Rockville, MD, USA), HuH7 cells were kindly supplied by J Gerin (Georgetown University Medical Center, Washington, DC, USA) and other urological tumor cell lines were kindly supplied by Dr Y Mizutani (Department of Urology, Kyoto University, Kyoto, Japan). E5 cells are ICP4 complementing cells derived from Vero 65 and were kindly supplied by N DeLuca (University of Pittsburgh School of Medicine, Pittsburgh, PA, USA). Viral stocks of wild-type HSV-1 strain KOS, obtained from D Knipe (Harvard Medical School, Boston, MA, USA), and ICP6…”

The UL4 gene of herpes simplex virus type 1 is dispensable for latency, reactivation and pathogenesis in mice.

“…The entire UL4 open reading frame (positions 12424-11827) is within this fragment. Plasmid pUL4HS was constructed by the insertion of a nonsense linker (DeLuca & Schaffer, 1987) containing stop codons in all three reading frames as well as a unique HpaI site into the XcmI site at position 12226 in the open reading frame of UL4. pUL4HS was cotransfected with infectious KOS DNA into Vero cells, progeny virus was harvested and plaques were screened by Southern blotting as described for an altered HpaI digestion pattern using $#P-labelled pUL4 as a probe.…”

Chronic lymphocytic leukemia B cells are highly sensitive to infection by herpes simplex virus-1 via herpesvirus-entry-mediator A

“…on E5 cells, a cell line derived from Vero cells that had been transfected with HSV-1 ICP4. 31,32 The MOI ratio provides the number of p.f.u. relative to the number of cells infected.…”