Specific inhibition of herpes virus replication by receptor-mediated entry of an antiviral peptide linked to Escherichia coli enterotoxin B subunit.
Mimetic peptides capable of selectively disrupting protein-protein interactions represent potential therapeutic agents for inhibition of viral and cellular enzymes. This approach was first suggested by the observation that the peptide YAGAVVNDL, corresponding to the carboxyl-terminal 9 amino acids of the small subunit of ribonucleotide reductase of herpes simplex virus, specifically inhibited the viral enzyme in vitro. Evaluation and use of this peptide as a potential antiviral agent has, however, been thwarted by its failure to inhibit virus replication in vivo, presumably because the peptide is too large to enter eukaryotic cells unaided. Here, we show that the nontoxic B subunit of Escherichia coli heat-labile enterotoxin can be used as a recombinant carrier for the receptor-mediated delivery of YAGAVVNDL into virally infected cells. The resultant fusion protein specifically inhibited herpes simplex virus type 1 replication and ribonucleotide reductase activity in quiescent Vero cells. Preincubation of the fusion protein with soluble GM1 ganglioside abolished this antiviral effect, indicating that receptor-mediated binding to the target cell is necessary for its activity. This provides direct evidence of the usefulness of carrier-mediated delivery to evaluate the intracellular efficacy of a putative antiviral peptide.
Marcello
A
A
Institute of Microbiology, University of Padova, Italy.
Loregian
A
A
Cross
A
A
Marsden
H
H
Hirst
T R
TR
Palù
G
G
D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents; D001427:Bacterial Toxins / Q000008:administration & dosage / Q000737:chemistry*; D001483:Base Sequence; D004768:Enterotoxins / Q000008:administration & dosage / Q000737:chemistry*; D029968:Escherichia coli Proteins; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011993:Recombinant Fusion Proteins; D012264:Ribonucleotide Reductases / Q000008:administration & dosage / Q000737:chemistry*; D018139:Simplexvirus / Q000254:growth & development*; D014709:Vero Cells; D014779:Virus Replication
D000998:Antiviral Agents; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; C027538:heat-labile enterotoxin, E coli; D012264:Ribonucleotide Reductases