The herpes simplex virus immediate early protein ICP27 encodes a potential metal binding domain and binds zinc in vitro.
The herpes simplex virus immediate early protein ICP27 encodes a potential metal binding domain and binds zinc in vitro.
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单纯疱疹病毒立即早期蛋白 ICP27 编码潜在的金属结合结构域并在体外结合锌。
DOI：
10.1016/0042-6822(92)90720-a
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发表时间：
1992
期刊：
影响因子：
3.7
通讯作者：
Sandri-Goldin,RM
中科院分区：
文献类型：
--
作者：
Vaughan,PJ;Thibault,KJ;Hardwicke,MA;Sandri-Goldin,RM
单纯疱疹病毒1型（HSV-1）立即早期调节蛋白ICP 27和ICP 0各自编码推定的锌指金属结合结构域。我们利用金属螯合亲和层析技术证明ICP 27和ICP 0能够在体外与锌结合。进一步利用该性质从HSV-1感染的细胞提取物中纯化ICP 27。ICP 27具有单链DNA结合活性。对体外翻译产生的ICP 27截短肽的分析证实，ICP 27的锌结合区位于羧基端105个氨基酸，跨越假定的金属结合基序。然而，正如移码突变以与野生型类似的效率结合的能力所证明的那样，该区域中半胱氨酸和组氨酸残基的特定配置并不需要发生结合。突变的肽保留四个组氨酸和半胱氨酸残基，但在不同的配置从共识提出的锌指基序。因此，虽然跨越ICP 27的金属结合结构域的区域对于激活子和阻遏子功能都是必需的，并且ICP 27在体外与锌结合，但尚不清楚体内锌结合是否是功能所必需的。
The herpes simplex virus type 1 (HSV-1) immediate-early regulatory proteins ICP27 and ICP0 each encode putative zinc-finger metal-binding domains. We utilized the technique of metal chelate affinity chromatography to demonstrate that ICP27 and ICP0 were able to bind to zincin vitro. This property was further exploited to purify ICP27 from extracts of HSV-1-infected cells. The purification procedure also revealed that ICP27 possessed single-stranded DNA-binding activity. Analysis of ICP27 truncated peptides produced byin vitrotranslation verified that the zinc-binding region of ICP27 resides in the carboxy terminal 105 amino acids spanning the putative metal binding motif. However, a specific configuration of cysteine and histidine residues in this region was not required for binding to occur as demonstrated by the ability of a frame-shift mutation to bind with an efficiency similar to wild type. The mutated peptide retained four histidine and cysteine residues but in a configuration different from the consensus proposed for zinc-finger motifs. Therefore, while the region spanning the metal binding domain of ICP27 is essential for both the activator and repressor functions, and ICP27 binds zincin vitro, it is not clear whether zinc bindingin vivois necessary for function.