The varicella-zoster virus immediate-early protein IE62 is a major component of virus particles.
Varicella-zoster virus (VZV) open reading frame (ORF) 62 potentially encodes a protein with considerable amino acid homology to the herpes simplex virus (HSV) immediate-early regulatory polypeptide ICP4 (or IE3). To identify and characterize its protein product(s) (IE62), we used a rabbit antiserum prepared against a synthetic peptide corresponding to the C-terminal 13 amino acids of the predicted protein. This antiserum reacted with phosphorylated polypeptides of 175 to 180 kDa that were made in VZV-infected cells and in cells infected with a vaccinia virus recombinant expressing IE62, but not in control-infected cells, confirming its specificity and reactivity to the IE62 protein. The antiserum recognized a 175-kDa polypeptide in purified virions that comigrated with a major structural protein. Comparison of this reactivity with that of an antipeptide antiserum directed against the VZV ORF 10 product (homologous to the HSV major structural protein VP16) indicates similar levels of ORF 62 and ORF 10 polypeptides in VZV virions. In contrast, antipeptide antiserum directed against the VZV ORF 29 product, the homolog of the HSV major DNA-binding protein, failed to recognize any protein in our virion preparations. Treatment of virions with detergents that disrupt the virion envelope did not dissociate IE62 from the nucleocapsid-tegument structure of the virion. Differential sensitivity of VZV virion IE62 to trypsin digestion in the presence or absence of Triton X-100 indicates that IE62 is protected from trypsin degradation by the virus envelope; since it is not a nucleocapsid protein, we conclude that it is part of the tegument. Finally, we show that the virion 175-kDa protein either can autophosphorylate or is a major substrate in vitro for virion-associated protein kinase activity.
Kinchington
P R
PR
Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-0499.
Hougland
J K
JK
Arvin
A M
AM
Ruyechan
W T
WT
Hay
J
J
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D014645:Herpesvirus 3, Human / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D017874:Immediate-Early Proteins; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010455:Peptides / Q000032:analysis; D010748:Phosphopeptides / Q000032:analysis; D054334:Viral Regulatory and Accessory Proteins / Q000032:analysis* / Q000235:genetics; D014771:Virion / Q000737:chemistry
D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D010455:Peptides; D010748:Phosphopeptides; D054334:Viral Regulatory and Accessory Proteins; C048925:herpes simplex virus, type 1 protein ICP4; D000255:Adenosine Triphosphate