Identification and characterization of a varicella-zoster virus DNA-binding protein by using antisera directed against a predicted synthetic oligopeptide.
We have identified, in varicella-zoster virus (VZV)-infected cells, the product of the gene predicted to code for the VZV analog of the herpes simplex virus major DNA-binding protein. The open reading frame of the VZV gene has the potential to code for a protein with a predicted molecular weight of 132,000 (a 132K protein). To detect the protein, a 12-amino-acid oligopeptide corresponding to the carboxyl terminus of the putative open reading frame was synthesized and used to prepare antisera in rabbits. The resulting antibodies reacted specifically in Western immunoblot analysis and immunoprecipitation with a single 130K polypeptide found in VZV-infected cells. The specific reactivity of the antisera with the 130K polypeptide was inhibited by the addition of synthetic peptide. Immunofluorescence studies with the antisera as probe for the 130K polypeptide suggested that this peptide is located predominantly within the nuclei of infected cells. Analysis of proteins that bind to single-stranded DNA immobilized on cellulose matrices indicated that 30 to 50% of the 130K polypeptide is capable of interacting with single-stranded DNA and that this interaction is overcome with 0.5 M NaCl. Thus, we have prepared a specific polyclonal antiserum that identifies a VZV DNA-binding protein whose properties are similar to those of the herpes simplex virus ICP8 (Vmw130) DNA-binding protein.
Kinchington
P R
PR
Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799.
Inchauspe
G
G
Subak-Sharpe
J H
JH
Robey
F
F
Hay
J
J
Ruyechan
W T
WT
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D003429:Cross Reactions; D003588:Cytopathogenic Effect, Viral; D004268:DNA-Binding Proteins / Q000276:immunology / Q000302:isolation & purification*; D005347:Fibroblasts; D014645:Herpesvirus 3, Human / Q000032:analysis* / Q000276:immunology; D007106:Immune Sera; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D011817:Rabbits; D018139:Simplexvirus / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology; D014779:Virus Replication
D004268:DNA-Binding Proteins; C045462:ICP8 protein, Simplexvirus; D007106:Immune Sera; D009842:Oligopeptides; D014764:Viral Proteins