Inhibition of Vaccinia virus entry by a broad spectrum antiviral peptide.
Concerns about the possible use of Variola virus, the causative agent of smallpox, as a weapon for bioterrorism have led to renewed efforts to identify new antivirals against orthopoxviruses. We identified a peptide, EB, which inhibited infection by Vaccinia virus with an EC(50) of 15 microM. A control peptide, EBX, identical in composition to EB but differing in sequence, was inactive (EC50>200 microM), indicating sequence specificity. The inhibition was reversed upon removal of the peptide, and EB treatment had no effect on the physical integrity of virus particles as determined by electron microscopy. Viral adsorption was unaffected by the presence of EB, and the addition of EB post-entry had no effect on viral titers or on early gene expression. The addition of EB post-adsorption resulted in the inhibition of beta-galactosidase expression from an early viral promoter with an EC(50) of 45 microM. A significant reduction in virus entry was detected in the presence of the peptide when the number of viral cores released into the cytoplasm was quantified. Electron microscopy indicated that 88% of the virions remained on the surface of cells in the presence of EB, compared to 37% in the control (p<0.001). EB also blocked fusion-from-within, suggesting that virus infection is inhibited at the fusion step. Analysis of EB derivatives suggested that peptide length may be important for the activity of EB. The EB peptide is, to our knowledge, the first known small molecule inhibitor of Vaccinia virus entry.
Altmann
S E
SE
Microbiology Doctoral Training Program, University of Wisconsin School of Medicine and Public Health, Madison, WI 53706, USA.
Jones
J C
JC
Schultz-Cherry
S
S
Brandt
C R
CR
D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D002459:Cell Fusion; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D010455:Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D013997:Time Factors; D014616:Vaccinia virus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D014771:Virion / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D053586:Virus Internalization / Q000187:drug effects*
D000998:Antiviral Agents; D010455:Peptides