Capsids are formed in a mutant virus blocked at the maturation site of the UL26 and UL26.5 open reading frames of herpes simplex virus type 1 but are not formed in a null mutant of UL38 (VP19C).
Previously we reported that null mutant viruses of UL19 (VP5) or of UL18 (VP23), essential components of herpes simplex virus type 1 (HSV-1) capsid shells, do not form precursor capsid structures as judged by sedimentation and electron microscope analysis. A goal of the present experiments was to isolate a null mutant virus for the remaining essential component of capsid shells, VP19C, encoded by the UL38 open reading frame (ORF). Furthermore, we wished to determine if a virus altered in the UL26 maturation cleavage site at residues 610 and 611 produced a lethal phenotype. Therefore, we decided to isolate cell lines that encode and express multiple capsid genes. Several cell lines were isolated by transformation of Vero cells and one designated C32 expressed all of the essential capsid proteins. Using this cell line we isolated a null mutant virus in the UL38 ORF and a mutant virus that was altered at residues 610 and 611 of the UL26 and UL26.5 gene products. We found that the null mutant in VP19C did not form a detectable product as judged by sedimentation and electron microscope analyses following infection of nonpermissive cells. The mutant virus altered at the UL26 maturation site resulted in the accumulation of B capsids. Therefore, cleavage at this site was essential for the maturation of B capsids into C capsids. Interestingly, the absence of cleavage at the maturation site was required for the retention of VP24 in the capsid.
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Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
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D000818:Animals; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics*; D002460:Cell Line; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000648:ultrastructure / Q000821:virology; D002522:Chlorocebus aethiops; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D016296:Mutagenesis; D016366:Open Reading Frames; D015183:Restriction Mapping; D012697:Serine Endopeptidases / Q000235:genetics*; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics*
C054582:ICP35 protein, Human herpesvirus 1; D014764:Viral Proteins; C079287:scaffold protein, Herpes simplex virus-1; D012697:Serine Endopeptidases