Herpes simplex virus VP16 forms a complex with the virion host shutoff protein vhs.
Herpes simplex virus (HSV) virions contain at least two regulatory proteins that modulate gene expression in infected cells: the transcriptional activator VP16 and the virion host shutoff protein vhs. VP16 stimulates transcription of the HSV immediate-early genes, and vhs suppresses host protein synthesis and induces accelerated turnover of cellular and viral mRNAs. We report here that vhs binds directly to VP16: vhs and VP16 were coprecipitated from infected cells by an anti-vhs antiserum, and vhs and VP16 protein A fusions each bound intact versions of the other protein in a solid-phase capture assay. In addition, vhs and VP16 interacted in the two-hybrid activator system when coexpressed in Saccharomyces cerevisiae. vhs residues 238 to 344 were sufficient for the interaction, and the VP16 acidic transcriptional activation domain was not required. vhs blocked the ability of VP16 to enter a multiprotein complex on an immediate-early TAATGARATTC consensus sequence, indicating that vhs interacts with one or more regions of VP16 required for promoter recognition. We suggest that this interaction may play a structural role in the assembly of HSV virions and modulate the activity of vhs during infection.
Smibert
C A
CA
Molecular Virology and Immunology Program, McMaster University, Hamilton, Ontario, Canada.
Popova
B
B
Xiao
P
P
Capone
J P
JP
Smiley
J R
JR
D000818:Animals; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000235:genetics; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics / Q000378:metabolism*; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012260:Ribonucleases; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013205:Staphylococcal Protein A / Q000096:biosynthesis / Q000235:genetics; D015533:Transcriptional Activation; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D017405:Herpes Simplex Virus Protein Vmw65; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D014764:Viral Proteins; C070722:virion host shutoff protein, Simplexvirus; D012260:Ribonucleases