Nucleotides within both proximal and distal parts of the consensus sequence are important for specific DNA recognition by the herpes simplex virus regulatory protein ICP4.
Nucleotides within both proximal and distal parts of the consensus sequence are important for specific DNA recognition by the herpes simplex virus regulatory protein ICP4.
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共有序列的近端和远端部分内的核苷酸对于单纯疱疹病毒调节蛋白 ICP4 的特异性 DNA 识别非常重要。
DOI：
10.1093/nar/19.3.477
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发表时间：
1991
影响因子：
14.9
通讯作者：
Litman,B
中科院分区：
文献类型：
--
作者：
Pizer,LI;Everett,RD;Tedder,DG;Elliott,M;Litman,B
单纯疱疹病毒 1 型调节蛋白 ICP4 是一种序列特异性 DNA 结合蛋白，与许多不同的位点相关，其中一些位点包括共有的 ATCGTCnnnnYCGRC。为了研究共有序列中保守碱基对 DNA 结合的参与，我们合成了一系列突变寡核苷酸并测试了它们与 ICP4 形成复合物的能力。我们还比较了细菌表达的 ICP4 片段（包括 DNA 结合域）的结合特异性。大多数（但不是全部）碱基的突变在共有序列的近端部分大大减少了 ICP4 的结合，影响远端部分的突变也是如此。甲基化干扰测定中鉴定出的大多数（但不是全部）G 残基都是有效结合所必需的。虽然细菌表达的包含氨基酸残基 252 – 523 的 ICP4 肽与 DNA 结合的特异性与整个蛋白质相似，但较短的蛋白质（残基 275 – 523）具有稍微宽松的 DNA 结合特异性。
The herpes simplex virus type 1 regulatory protein ICP4 is a sequence specific DNA binding protein which associates with a number of different sites, some of which include the consensus ATCGTCnnnnYCGRC. In order to investigate the involvement in DNA binding of conserved bases within the consensus, we have synthesised a family of mutant oligonucleotides and tested their ability to form a complex with ICP4. We have also compared the binding specificities of bacterially expressed fragments of ICP4 which include the DNA binding domain. Mutation of most (but not all) bases In the proximal part of the consensus greatly reduced binding by ICP4, as did a mutation affecting the distal part. Most (but not all) G residues identified in methylatlon interference assays were required for efficient binding. While a bacterially expressed ICP4 peptide encompassing amino acid residues 252 – 523 bound to DNA with a specificity similar to that of the whole protein, a shorter protein (residues 275 – 523) had a slightly relaxed DNA binding specificity.