Mutation of a single lysine residue severely impairs the DNA recognition and regulatory functions of the VZV gene 62 transactivator protein.
The product of varicella-zoster virus gene 62 (VZV 140k) is a potent transactivator protein. We have identified a region within the DNA binding domain of VZV 140k that shows a striking similarity to the DNA recognition helix of the homeodomain, with an especially highly conserved quartet of residues, WLQN. The 140k protein has functional counterparts within the other alphaherpesviruses, which include the major transcriptional regulatory protein of HSV-1, (ICP4), and the WLQN region is highly conserved among the members of this family of viral transactivators. Substitution of VZV 140k residue lysine 548, just adjacent to the WLQN region, drastically reduces the DNA binding activity of the 140k DNA binding domain and the intact 140k mutant protein fails to activate gene expression. Substitutions of two other VZV 140k residues in this conserved WLQN region result in alterations to the DNA binding interaction and reduced transactivation activities. All three mutations act at the level of DNA recognition, as they have no apparent effect on the dimerization state, solubility or efficiency of expression of the mutant peptides.
Tyler
J K
JK
MRC Virology Unit, Glasgow, UK.
Allen
K E
KE
Everett
R D
RD
D000595:Amino Acid Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry*; D005801:Genes, Homeobox; D014645:Herpesvirus 3, Human / Q000235:genetics*; D017874:Immediate-Early Proteins / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D015534:Trans-Activators / Q000737:chemistry*; D014759:Viral Envelope Proteins / Q000737:chemistry*
D004268:DNA-Binding Proteins; C074221:IE62 protein, Human herpesvirus 3; D017874:Immediate-Early Proteins; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D008239:Lysine