title: A mutation in UL15 of herpes simplex virus 1 that reduces packaging of cleaved genomes.
abstract: Herpesvirus genomic DNA is cleaved from concatemers that accumulate in infected cell nuclei. Genomic DNA is inserted into preassembled capsids through a unique portal vertex. Extensive analyses of viral mutants have indicated that intact capsids, the portal vertex, and all components of a tripartite terminase enzyme are required to both cleave and package viral DNA, suggesting that DNA cleavage and packaging are inextricably linked. Because the processes have not been functionally separable, it has been difficult to parse the roles of individual proteins in the DNA cleavage/packaging reaction. In the present study, a virus bearing the deletion of codons 400 to 420 of U(L)15, encoding a terminase component, was analyzed. This virus, designated vJB27, failed to replicate on noncomplementing cells but cleaved concatemeric DNA to ca. 35 to 98% of wild-type levels. No DNA cleavage was detected in cells infected with a U(L)15-null virus or a virus lacking U(L)15 codons 383 to 385, comprising a motif proposed to couple ATP hydrolysis to DNA translocation. The amount of vJB27 DNA protected from DNase I digestion was reduced compared to the wild-type virus by 6.5- to 200-fold, depending on the DNA fragment analyzed, thus indicating a profound defect in DNA packaging. Capsids containing viral DNA were not detected in vJB27-infected cells, as determined by electron microscopy. These data suggest that pU(L)15 plays an essential role in DNA translocation into the capsid and indicate that this function is separable from its role in DNA cleavage.
authors: Department of Microbiology and Immunology, C5143 Veterinary Education Center, Cornell University, Ithaca, NY 14853, USA.
authors: Kui
authors: K
authors: Yang
authors: Elizabeth G
authors: EG
authors: Wills
authors: Joel D
authors: JD
authors: Baines
mesh_terms: D000818:Animals; D002213:Capsid / Q000378:metabolism; D002460:Cell Line; D004279:DNA, Viral / Q000378:metabolism*; D005816:Genetic Complementation Test; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D004279:DNA, Viral; D050505:Mutant Proteins; D014764:Viral Proteins; C069185:DNA cleavage and packaging proteins, Herpesvirus