Physical interaction between two varicella zoster virus gene regulatory proteins, IE4 and IE62.
Transfection assays demonstrate that the varicella zoster virus (VZV) immediate-early 62 (IE62) protein is a major transactivator of VZV gene expression, whereas a second immediate-early protein, IE4, can act as a major coactivator of transactivation mediated through IE62. To test whether IE62 and IE4 interact physically, we performed several protein-protein interaction assays. Coimmunoprecipitation analyses using VZV-infected cell lysates as well as purified protein mixtures demonstrate that IE62 and IE4 form stable complexes in solution under stringent salt conditions. Enzyme-linked immunosorbent assay protein-protein interaction assays and maltose-binding protein capture assays demonstrate that IE62 binds IE4 in a concentration- and dose-dependent manner. Far Western blot analyses show that IE4 binds to an undermodified form of IE62, and the use of calf intestinal phosphatase and protein kinases suggests that the interaction with IE4 is dependent on the phosphorylation state of IE62. An IE4 binding domain on IE62 has been mapped using a set of truncated IE62 fusion peptides. Collectively, these results imply a direct and specific physical interaction between IE4 and less-phosphorylated forms of IE62. These data have implications for virion assembly, as well as for the regulation of gene expression in VZV-infected cells.
Spengler
M L
ML
Department of Microbiology and Markey Center for Microbial Pathogenesis, State University of New York at Buffalo School of Medicine, Buffalo, New York, 14214, USA.
Ruyechan
W T
WT
Hay
J
J
D000595:Amino Acid Sequence; D001665:Binding Sites; D015967:Gene Expression Regulation, Viral; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010766:Phosphorylation; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins
C074221:IE62 protein, Human herpesvirus 3; D017874:Immediate-Early Proteins; C085824:ORF4 protein, Human herpesvirus 3; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D014764:Viral Proteins