Binding of the herpes simplex virus type 1 UL9 gene product to an origin of viral DNA replication.

Binding of the herpes simplex virus type 1 UL9 gene product to an origin of viral DNA replication.
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1 型单纯疱疹病毒 UL9 基因产物与病毒 DNA 复制起点的结合。

DOI:
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发表时间:
1989
影响因子:
14.9
通讯作者:
N. Stow
N. Stow
中科院分区:
生物学2区
文献类型:
--
作者:
H. M. Weir;Janice M. Calder;N. Stow

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1 型单纯疱疹病毒 (HSV-1) 编码的多肽与病毒 DNA 复制起点的结合已通过凝胶阻滞测定进行了研究。将 HSV-1 感染细胞的核提取物与标记的含起点片段一起孵育,导致形成特定的延迟复合物,在与 UL9 基因产物反应的抗体存在下,该复合物的迁移进一步减少。在立即早期 (IE) 启动子的控制下引入额外的 UL9 基因拷贝,赋予了在感染 IE 阶段被阻断的 HSV-1 ts 突变体在不允许的温度下表达起源结合活性的能力。内源或外源蛋白水解活性揭示了相对蛋白酶抗性结构域的存在,该结构域保留了序列特异性DNA结合活性。在大肠杆菌中表达为融合蛋白的UL9基因的C端317个氨基酸也与起点结合。我们的结果表明,UL9 基因产物与病毒起源结合,并且序列特异性识别和结合由多肽的 C 端 37% 指定。
The binding of a herpes simplex virus type 1 (HSV-1) encoded polypeptide to a viral origin of DNA replication has been studied by using a gel retardation assay. Incubation of nuclear extract from HSV-1 infected cells with a labelled origin-containing fragment resulted in the formation of a specific retarded complex, the migration of which was further reduced in the presence of an antibody reactive with the UL9 gene product. Introduction of an additional copy of the UL9 gene, under the control of an immediate early (IE) promoter, conferred the ability to express origin binding activity at the non-permissive temperature upon an HSV-1 ts mutant blocked at the IE stage of infection. Endogenous or exogenous proteolytic activity revealed the presence of a relatively protease-resistant domain which retained sequence-specific DNA binding activity. The C-terminal 317 amino acids of the UL9 gene expressed as a fusion protein in Escherichia coli also bound to the origin. Our results demonstrate that the UL9 gene product binds to a viral origin and that sequence specific recognition and binding are specified by the C-terminal 37% of the polypeptide.
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