title: Complementation of a gl-deficient feline herpesvirus recombinant by allotopic expression of truncated gl derivatives.
abstract: The alphaherpesvirus glycoproteins gE and gI form a hetero-oligomeric complex involved in cell-to-cell transmission. The gI-deficient recombinant feline herpesvirus (FHV), FHVdeltagI-LZ, produces plaques that are only 15% the size of those of wild-type FHV. Here, we have complemented FHV(delta)gI-LZ allotopically by expressing intact gI and C-terminally truncated gI derivatives from the thymidine kinase locus. The effect on gE-gI-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 gI fully restored plaque size. Deletion of the C-terminal 11 residues of gI did not affect cell-to-cell spread, whereas deletion of the complete cytoplasmic tail reduced plaque size by only 35%. Mutants expressing gI166, 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 gI-deficient controls, indicating that the gE-gI166 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-gI function in vitro.
authors.forename: J D
authors.initials: JD
authors.lastname: Mijnes
authors.forename: C
authors.initials: C
authors.lastname: Vlot
authors.forename: J B
authors.initials: JB
authors.lastname: Buntjer
authors.forename: J
authors.initials: J
authors.lastname: van den Broek
authors.forename: M C
authors.initials: MC
authors.lastname: Horzinek
authors.forename: P J
authors.initials: PJ
authors.lastname: Rottier
authors.forename: R J
authors.initials: RJ
authors.lastname: de Groot
mesh_terms: D018138:Alphaherpesvirinae / Q000235:genetics* / Q000472:pathogenicity; D000818:Animals; D002415:Cats; D004274:DNA, Recombinant; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D014759:Viral Envelope Proteins / Q000235:genetics*; D014774:Virulence / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
chemical_list: D004274:DNA, Recombinant; D014759:Viral Envelope Proteins; C077407:glycoprotein L, Human herpesvirus 1