Arbuckle M I, Stow N D
MRC Virology Unit, Institute of Virology, Glasgow, U.K.
J Gen Virol. 1993 Jul;74 ( Pt 7):1349-55. doi: 10.1099/0022-1317-74-7-1349.
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.
单纯疱疹病毒1型的起始结合蛋白由UL9基因编码。我们之前描述过一种质粒pB1，它编码一种融合蛋白，该融合蛋白仅包含UL9多肽的C末端317个氨基酸，并表明该产物保留了序列特异性DNA结合能力。现在构建了两个系列的pB1突变体，并对这些多肽进行了起始结合活性测试。通过C末端截短，我们发现UL9的C末端34个氨基酸对于结合是可有可无的，而必需残基位于801至818位之间。对一系列在不同位置插入四个氨基酸的突变体进行分析，确定了DNA结合结构域中改变要么消除结合活性要么对结合活性影响相对较小的区域。两个结合活性处于中间水平的突变体也表现出温度或序列特异性效应。
