Mouse Anti-FHV-1 monoclonal antibody, clone IN650

To identify new genes affecting Tomato bushy stunt virus (TBSV) replication in yeast model host, we are studying protein families, whose members have been identified during previous high throughput screening. In this paper, we have characterized the WW domain-containing protein family from yeast and plants. We find that, in addition to Rsp5 E3 ubiquitin ligase, yeast Wwm1 and Prp40 and three Arabidopsis WW domain-containing proteins are strong inhibitors of TBSV replication. The tombusvirus replicase complex isolated from yeast with down-regulated Wwm1 protein level was more active. Accumulation of viral p92(pol) was reduced when Wwm1 was over-expressed, suggesting that the stability of p921(pol) might be reduced, as observed with Rsp5. Moreover, replication of two insect RNA viruses is also inhibited by Wwm1 and Rsp5, suggesting that WW domain-containing proteins might have broad regulatory effects on RNA viruses. Thus, artificial antiviral proteins with WW domains could be useful antiviral strategy. (C) 2012 Published by Elsevier Inc.

The alphaherpesvirus glycoproteins gE and gl farm a hetero-oligomeric complex involved in cell-to-cell transmission. The gl-deficient recombinant feline herpesvirus (FHV), FHV Delta gl-LZ, produces plaques that are only 15% the size of those of wild-type FHV. Here, we have complemented FHV Delta gl-LZ allotopically by expressing intact gl and C-terminally truncated gl derivatives from the thymidine kinase locus. The effect on gE-gl-mediated cell-to-cell spread was assessed by plaque assay employing computer-assisted image analysis (software available at http://www.androclus.vet.uu.nl/ spotter/spotter.htm). Allotopic complementation with intact gl fully restored plaque size. Deletion of the C-terminal 11 residues of gl did not affect cell-to-cell spread, whereas deletion of the complete cytoplasmic tail reduced plaque size by only 35%. Mutants expressing gl(166), roughly corresponding to the N-terminal half of the ectodomain, displayed a small-plaque phenotype. Nevertheless, their plaques were reproducibly larger than those of matched gl-deficient controls, indicating that the gE-gl(166) hetero-oligomer, though crippled, is still able to mediate cell-to-cell spread. Our data demonstrate that plaque analysis provides a reliable and convenient tool to measure and quantitate gE-gl function in vitro.