title: The null mutant of the U(L)31 gene of herpes simplex virus 1: construction and phenotype in infected cells.
abstract: Earlier studies have shown that the U(L)31 protein is homogeneously distributed throughout the nucleus and cofractionates with nuclear matrix. We report the construction from an appropriate cosmid library a deletion mutant which replicates in rabbit skin cells carrying the U(L)31 gene under a late (gamma1) viral promoter. The mutant virus exhibits cytopathic effects and yields 0.01 to 0.1% of the yield of wild-type parent virus in noncomplementing cells but amounts of virus 10- to 1,000-fold higher than those recovered from the same cells 3 h after infection. Electron microscopic studies indicate the presence of small numbers of full capsids but a lack of enveloped virions. Viral DNA extracted from the cytoplasm of infected cells exhibits free termini indicating cleavage/packaging of viral DNA from concatemers for packaging into virions, but analyses of viral DNAs by pulsed-field electrophoresis indicate that at 16 h after infection, both the yields of viral DNA and cleavage of viral DNA for packaging are decreased. The repaired virus cannot be differentiated from the wild-type parent. These results suggest the possibility that U(L)31 protein forms a network to enable the anchorage of viral products for the synthesis and/or packaging of viral DNA into virions.
authors.affiliation: The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, Illinois 60637, USA.
authors.forename: Y E
authors.initials: YE
authors.lastname: Chang
authors.forename: C
authors.initials: C
authors.lastname: Van Sant
authors.forename: P W
authors.initials: PW
authors.lastname: Krug
authors.forename: A E
authors.initials: AE
authors.lastname: Sears
authors.forename: B
authors.initials: B
authors.lastname: Roizman
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002213:Capsid / Q000648:ultrastructure; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000378:metabolism*; D003673:Defective Viruses / Q000235:genetics / Q000254:growth & development; D017353:Gene Deletion; D005814:Genes, Viral; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000235:genetics / Q000254:growth & development*; D006801:Humans; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D009687:Nuclear Proteins / Q000378:metabolism / Q000502:physiology*; D011817:Rabbits; D014709:Vero Cells; D014764:Viral Proteins / Q000502:physiology*; D015678:Viral Structural Proteins / Q000235:genetics; D014771:Virion / Q000648:ultrastructure; D014779:Virus Replication
chemical_list: D004279:DNA, Viral; D009687:Nuclear Proteins; C108721:UL31 protein, Human herpesvirus 1; D014764:Viral Proteins; D015678:Viral Structural Proteins