Characterization of ICP6::lacZ insertion mutants of the UL15 gene of herpes simplex virus type 1 reveals the translation of two proteins.
The herpes simplex virus type 1 (HSV-1) UL15 gene is a spliced gene composed of two exons and is predicted to encode an 81-kDa protein of 735 amino acids (aa). Two UL15 gene products with molecular masses of 75 and 35 kDa have been observed (J. Baines, A. Poon, J. Rovnak, and B. Roizman, J. Virol. 68:8118-8124, 1994); however, it is not clear whether the smaller form represents a proteolytic cleavage product of the larger form or whether it is separately translated. In addition, an HSV-1 temperature-sensitive mutant in the UL15 gene (ts66.4) is defective in both cleavage of viral DNA concatemers into unit-length monomers and packaging of viral DNA into capsids (A. Poon and B. Roizman, J. Virol. 67:4497-4503, 1993; J. Baines et al., J. Virol. 68:8118-8124, 1994). In this study, we detected two UL15 gene products of 81 and 30 kDa in HSV-1-infected cells, using a polyclonal antibody raised against a maltose binding protein fusion construct containing UL15 exon 2. In addition, we report the isolation of two HSV-1 insertion mutants, hr81-1 and hr81-2, which contain an ICP6::lacZ insertion in UL15 exon 1 and exon 2 and thus would be predicted to encode C-terminally truncated peptides of 153 and 509 aa long, respectively. hr81-1 and hr81-2 are defective in DNA cleavage and packaging and accumulate only B capsids. However, both mutants are able to undergo wild-type levels of DNA replication and genomic inversion, suggesting that genomic inversion is a result of DNA replication rather than of DNA cleavage and packaging. We also provide evidence that the 81- and 30-kDa proteins are the products of separate in-frame translation events from the UL15 gene and that the 81-kDa full-length UL15 protein is required for DNA cleavage and packaging.
Yu
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D
Department of Microbiology, University of Connecticut Health Center, Farmington 06030, USA.
Sheaffer
A K
AK
Tenney
D J
DJ
Weller
S K
SK
D000818:Animals; D002213:Capsid / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D007446:Chromosome Inversion; D004279:DNA, Viral / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007763:Lac Operon; D016254:Mutagenesis, Insertional; D010641:Phenotype; D014176:Protein Biosynthesis; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics*; D019065:Virus Assembly
D004279:DNA, Viral; D014764:Viral Proteins; C054366:herpes simplex virus type 1-protein ICP6; C069185:DNA cleavage and packaging proteins, Herpesvirus