Proteins Specified by Herpes Simplex Virus XII. The Virion Polypeptides of Type 1 Strains

Abstract

The polypeptides from purified virions of a herpes simplex 1 (human herpes-virus 1) strain, F1, which had been passaged a limited number of times in cell culture after isolation, formed 33 bands on electrophoretic separation in polyacrylamide gels cross-linked with N, N′ -diallyltartardiamide in contrast to a maximum resolution of only 24 to 25 bands in gels cross-linked with N, N′ -methylenebisacrylamide. This increase in the number of bands was due chiefly to an improved separation of glycosylated polypeptides from nonglycosylated polypeptides with which they co-electrophoresed on methylenebisacrylamide cross-linked gels. Purified virions of HSV-1 [F1] had a protein/DNA mass ratio of 10.7 ± 0.96, and based on a DNA molecular mass of 85 × 10 6 to 100 × 10 6 the estimated weight of virion polypeptides ranges from 16.4 to 19.4 × 10 −16 g. The number of molecules of each polypeptide per virion ranged from less than 50 to 1,500. Comparison of the virion polypeptides of two HSV-1 strains with similar isolation and limited passage history with those of four HSV-1 strains with histories of numerous passages outside the human host showed a number of nonrandom variations in virion polypeptides. Thus, although the virion polypeptides of two strains with similar isolation and limited passage history could not be differentiated, strains with extended passage histories differed markedly from each other and from the limited passage strains in the number and electrophoretic mobility of noncapsid polypeptides and notably in those of the envelope.

O-linked oligosaccharides are acquired by herpes simplex virus glycoproteins in the Golgi apparatus

“…were purified by preparative electrophoresis on 1.5 mm thick, 8.5% polyacrylamide gels crosslinked with N,N'-Diallyltartardiamide (DATD) (Heine et al, 1974). SDS extracts of 'C-glucosamine-or "C-galactose-labeled cells were sonicated and boiled for 5 min before being electrophoresed for 4.5 hr at 15 mA per gel.…”

“…Alternatively, HSVl glycoproteins were isolated by immunoprecipitation with monoclonal antibodies (l-144, specific for gB; 11-474, specific for gC; 11-436. specific for gD; or 11-481, specific for gE) as previously described (Johnson and Spear, 1982) and dialyzed against 0.1% SDS for treatment with enzymes or eluted into 2% SDS, 50 mM Tris-HCI (pH 6.8). 10% glycerol for electrophoresis on 8.5% polyacrylamide gels crosslinked with DATD (Heine et al, 1974). Analytical gels were impregnated with 2.5-diphenyloxazole (Banner and Laskey, 1974) and then dried and placed in contact with Cronex Medical x-ray film at -70°C.…”

Bacterial artificial chromosome derived simian varicella virus is pathogenic in vivo

“…SVV and VZV ORF22 are putative tegument proteins based on homology to HSV-1 UL36 [48]. UL36 is a HSV-1 late gene and the HSV-1 virion contains 100-150 copies of UL36 [49,50]. UL36 is essential to HSV-1 replication and the phenotype of a null mutant virus showed accumulation of capsids containing cytosolic DNA that did not mature into enveloped virions [51][52][53].…”

The herpes simplex virus type 1 particle: structure and molecular functions

“…Heine rt ul. (86) estimated that there would be less than 150 molecules per particle. From mapping studies using intertypic recombinants (143,162), analysis of a temperature-sensitive mutant (8) and examination of the sequence of the HSV-1 genome, McGeoch et 01.…”