A mutational analysis of the DNA-binding domain of the herpes simplex virus type 1 UL9 protein

Abstract

The herpes simplex virus type 1 origin-binding protein is encoded by gene UL9. We previously described a plasmid, pB1, which encodes a fusion protein containing only the C-terminal 317 amino acids of the UL9 polypeptide and showed that this product retains sequence-specific DNA-binding ability. Two series of pB1 mutants have now been constructed and the polypeptides were tested for origin-binding activity. Using C-terminal truncations, we show that the C-terminal 34 amino acids of UL9 are dispensable for binding and that essential residues lie between positions 801 and 818. Analysis of a series of mutants containing insertions of four amino acids at various positions identified regions of the DNA-binding domain in which alterations either abolished or had relatively little effect upon binding activity. Two mutants which were intermediate in their binding activities also exhibited temperature- or sequence-specific effects.