Packaging of genomic and amplicon DNA by the herpes simplex virus type 1 UL25-null mutant KUL25NS.
The herpes simplex virus type 1 (HSV-1) mutant KUL25NS, containing a null mutation within the UL25 gene, was isolated and characterized by McNab and coworkers (A. R. McNab, P. Desai, S. Person, L. L. Roof, D. R. Thomsen, W. W. Newcomb, J. C. Brown, and F. L. Homa, J. Virol. 72:1060-1070, 1998). This mutant was able to cleave the concatemeric products of viral DNA replication into monomeric units, but in contrast to wild-type (wt) HSV-1, they were degraded by DNase treatment, indicating that they were not stably packaged into virus capsids. I have examined the packaging of the KUL25NS genome and an HSV-1 amplicon in cells infected with the mutant virus. In contrast to the previous results, a low level of KUL25NS DNA was resistant to DNase digestion, indicating that it was retained in capsids. The proportion of this packaged DNA present as full-length genomes was much lower than in cells infected by wt HSV-1, and there was a significant overrepresentation of the long terminus and underrepresentation of the short terminus. KUL25NS was less impaired in stably packaging amplicon DNA than in packaging its own genome, and the packaged molecules contained approximately equimolar amounts of the two terminal fragments. Below about 100 kbp, the packaged amplicon molecules exhibited an abundance and size distribution similar to those generated using wt HSV-1 as a helper, but the mutant was relatively impaired in packaging longer amplicon molecules. Both packaged genomic and amplicon DNAs were retained in the nuclei of KUL25NS-infected cells. These results suggest that the UL25 protein may play an important role during the later stages of the head-filling process, prior to release of capsids into the cytoplasm.
Stow
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ND
MRC Virology Unit, Institute of Virology, Glasgow G11 5JR, United Kingdom. n.stow@vir.gla.ac.uk
D000818:Animals; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004279:DNA, Viral / Q000032:analysis / Q000502:physiology*; D016521:Electrophoresis, Gel, Pulsed-Field; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D009154:Mutation; D014709:Vero Cells; D014764:Viral Proteins / Q000502:physiology*; D019065:Virus Assembly
D004279:DNA, Viral; D014764:Viral Proteins