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        Inhibition of Herpes Simplex Virus Type 1 by Human Salivary Cystatin SN and its Proteinase Inhibitory Regions

        Hiltke, Tara R.,Kennell, Alan F.,Bergey, Earl J.,Bobek, Libuse A. Korean Academy of Oral Biology and the UCLA Dental 2000 International Journal of Oral Biology Vol.25 No.1

        Cystatins are a family cysteine proteinase inhibitors, many of which possess antiviral properties. Previous studies demonstrated that human cystatin C and salivary cystatin SN inhibit herpes simplex virus type 1〔HSV-1〕replication. The full-length cystatin C and SN do not function as serine proteinase inhibitors, but contain serine proteinase inhibitory regions. The goal of this study was to evaluate whether the inhibition of HSV-1 by cystatin SN occurs through its cysteine or serine proteinase inhibitory regions. using viral yield reduction assays, our results demonstrated that the N-truncated cystatin SN〔a deletion of one of the conserved cysteine proteinase inhibitory region〕was not able to effectively inhibit HSV-1〔20% versus 90% by the full-length cystatin SN〕. The results also showed that a circular peptide to the serine proteinase inhibitory region of cystatin SN inhibited HSV-1 by only 20%, while the linear peptide to the same region failed to inhibit HSV-1. Previous studies indicated that the inhibition of HSV-1 replicatin by cystatin SN occurred during the later stages of the viral infection. Thus, we have examined, by western blot analysis, the effect of cystatin SN on specific HSV-1 structural proteins 〔VP5, VP19, VP23, VP21/VP22a〕, as well as its effect on a non-structural DNA binding protein produced at an earlier stage in the viral life cycle〔ICP8〕. We report that cystatin SN reduced the production of total HSV-1 proteins, as well as all the individual proteins tested, while the circular peptide inhibited only two of the proteins tested, VP5 and VP19. In conclusion, the role of cystatin SN in HSV-1 inhibition appears to involve both ist cysteine and serine proteinase inhibitory regions.

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