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

• Published in: Journal of general virology 1993-07, Vol.74 (7), p.1349-1355
• Main Authors: Arbuckle, Margaret I, Stow, Nigel D
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Base Sequence; Binding Sites; Biological and medical sciences; Blotting, Western; Cloning, Molecular; DNA Mutational Analysis; DNA-Binding Proteins - biosynthesis; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genetics; herpes simplex virus 1; Microbiology; Molecular Sequence Data; Molecular Weight; Mutagenesis, Insertional; Oligodeoxyribonucleotides; Plasmids; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Restriction Mapping; Sequence Deletion; Simplexvirus - genetics; Simplexvirus - metabolism; Viral Proteins - biosynthesis; Viral Proteins - genetics; Viral Proteins - metabolism; Virology
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-74-7-1349

MRC Virology Unit, Institute of Virology, Church Street, Glasgow G11 5JR, U.K. 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. Received 14 December 1992; accepted 15 February 1993.