The null mutant of the U(L)31 gene of herpes simplex virus 1: construction and phenotype in infected cells.
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.
Chang
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YE
The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, Illinois 60637, USA.
Van Sant
C
C
Krug
P W
PW
Sears
A E
AE
Roizman
B
B
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
D004279:DNA, Viral; D009687:Nuclear Proteins; C108721:UL31 protein, Human herpesvirus 1; D014764:Viral Proteins; D015678:Viral Structural Proteins