The DNA binding domains of the varicella-zoster virus gene 62 and herpes simplex virus type 1 ICP4 transactivator proteins heterodimerize and bind to DNA.
The product of varicella-zoster virus gene 62 (VZV 140k) is the functional counterpart of the major transcriptional regulatory protein of herpes simplex virus type 1 (HSV-1), ICP4. We have found that the purified bacterially expressed DNA binding domain of VZV 140k (residues 417-647) is a stable dimer in solution. As demonstrated by the appearance of a novel protein--DNA complex of intermediate mobility in gel retardation assays, following in vitro co-translation of a pair of differently sized VZV 140k DNA binding domain peptides, the 140k DNA binding domain peptide binds to DNA as a dimer. In addition, the DNA binding domain peptide of HSV-1 ICP4 readily heterodimerizes with the VZV 140k peptide on co-translation, indicating that HSV-1 ICP4 and VZV 140k possess very similar dimerization interfaces. It appears that only one fully wild type subunit of the dimer is sufficient to mediate sequence specific DNA recognition in certain circumstances. Co-immunoprecipitation analysis of mutant DNA binding domain peptides, co-translated with an epitope-tagged ICP4 DNA binding domain, shows that the sequence requirements for dimerization are lower than those necessary for DNA binding.
Tyler
J K
JK
MRC Virology Unit, Glasgow, UK.
Everett
R D
RD
D001665:Binding Sites; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000378:metabolism*; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000378:metabolism*; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014176:Protein Biosynthesis; D012996:Solutions; D015534:Trans-Activators / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C074221:IE62 protein, Human herpesvirus 3; D017874:Immediate-Early Proteins; D012996:Solutions; D015534:Trans-Activators; D014157:Transcription Factors; D014759:Viral Envelope Proteins; C048925:herpes simplex virus, type 1 protein ICP4