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. 1993 Feb 11;21(3):513–522. doi: 10.1093/nar/21.3.513

The DNA binding domain of the varicella-zoster virus gene 62 protein interacts with multiple sequences which are similar to the binding site of the related protein of herpes simplex virus type 1.

J K Tyler 1, R D Everett 1
PMCID: PMC309147  PMID: 8382799

Abstract

Varicella-zoster virus gene 62 encodes a protein with predicted Mr of 140,000D (VZV 140k) that shares extensive predicted amino acid sequence homology with the major immediate early (IE) transcriptional regulator protein of herpes simplex virus type 1 (HSV-1) Vmw175. The integrity of highly conserved region 2 is essential for the DNA binding and transcriptional regulatory functions of Vmw175. Similarly, an insertion mutation in region 2 (codons 468-641) of 140k eliminates the transcriptional repression and activation functions of this protein. We have expressed a fragment of 140k which encompasses region 2 as a non-fusion polypeptide in bacteria. This 140k DNA binding domain peptide (codons 417-646) binds to numerous DNA sequences throughout the VZV gene 62 promoter region. It induces multiple regions of protection from DNase I digestion, flanked by sites of DNase I hypersensitivity. Several of the sites recognized can be considered to be divergent forms of the consensus sequence which is recognized by Vmw175. However, by use of a panel of mutagenized probe fragments, we found that the 140k DNA binding domain was less sequence-specific than Vmw175 in its interactions with DNA. Consistent with this, the homologous Vmw175 DNA binding domain, and also intact Vmw175, recognize the gene 62 binding sites much less efficiently than the 140k DNA binding domain. Also in contrast to the situation with Vmw175, the 140k DNA binding domain failed to induce DNA bending when occupying the binding sites in its own promoter. Deletion analysis has mapped the minimal DNA binding domain of the VZV 140k protein, as measured in gel retardation analysis, to lie within residues 472 to 633. The differences in binding characteristics of the DNA binding domains of the homologous VZV 140k and HSV-1 Vmw175 IE proteins may account for the subtle differences in their regulatory activities in transfection assays and during virus growth in tissue culture.

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Selected References

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