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首页> 外文期刊>Virology >AP-1 CIS-RESPONSE ELEMENTS ARE INVOLVED IN BASAL EXPRESSION AND VMW 110 TRANSACTIVATION OF THE LARGE SUBUNIT OF HERPES SIMPLEX VIRUS TYPE 2 RIBONUCLEOTIDE REDUCTASE (ICP10)
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AP-1 CIS-RESPONSE ELEMENTS ARE INVOLVED IN BASAL EXPRESSION AND VMW 110 TRANSACTIVATION OF THE LARGE SUBUNIT OF HERPES SIMPLEX VIRUS TYPE 2 RIBONUCLEOTIDE REDUCTASE (ICP10)

机译：AP-1 CIS反应元素涉及基础表达和单纯疱疹病毒2型核糖核苷酸还原酶（ICP10）的大亚基的VMW 110交易

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摘要

The promoter of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) has two AP-I cis-response elements, respectively located at positions -62 and -94 relative to the transcription start site (Wymer et al., 1989, J. Virol. 63, 2773-2784). Chloramphenicol acetyl transferase (CAT) analysis with hybrid constructions of the CAT structural gene and the ICP10 promoter or its mutants and gel retardation studies were used to examine the role of the AP-1 cis-response elements in expression from the ICP10 promoter. Basal expression from the wild-type promoter was significantly (75-90%) reduced by mutation of the upstream or downstream AP-1 element. Mutation in the upstream AP-1 element also caused a 60% reduction in c-Jun-mediated activation. Activation was decreased 40% by mutation in the downstream AP-1 element and it was abrogated by mutation of both elements. Similar results were obtained for ACT-deleted mutants and mutants in which CT was mutated to AG. The Irans-activation by Vmw110 was also reduced by mutation of the AP-1 elements (10- and 2-fold for the upstream and downstream element, respectively) and it was abrogated by mutation of both AP-1 elements. Mutation of nucleotides adjacent to the AP-1 cis-response elements had no effect on trans-activation. Gel retardation assays with a DNA probe representing the wild-type ICP10 promoter and nuclear extracts from HSV-1-infected cells identified one complex that was not seen with mock-infected cells or with cells infected with a Vmw110-deleted mutant. The complex was not seen when HSV-1-infected cells were reacted with an AP-1-mutant DNA probe, and its formation was competed by an AP-1 but not a mutant AP-1 oligonucleotide. The migration of this complex was retarded by c-Fos antibody, suggesting that both AP-1 and Vmw110 are involved in its formation. A mutant deleted in all sequences upstream of the TATA box was also activated by Vmw110, but this activation was only 2-fold lower than that seen for the wild type and significantly higher (10-fold) than that seen for the double AP-1 mutants. The data suggest that AP-1 elements play a crucial role in ICP10 gene expression/activation.
机译：单纯疱疹病毒2型核糖核苷酸还原酶（ICP10）大亚基的启动子具有两个AP-1顺式反应元件，分别位于相对于转录起始位点的-62和-94位置（Wymer等，1989; J.Virol.63，2773-2784）。氯霉素乙酰基转移酶（CAT）分析与CAT结构基因和ICP10启动子或其突变体的杂合构建以及凝胶阻滞研究用于检查AP-1顺式反应元件在ICP10启动子表达中的作用。通过上游或下游AP-1元件的突变，野生型启动子的基础表达显着降低（75-90％）。上游AP-1元件的突变也导致c-Jun介导的激活减少了60％。下游AP-1元件的突变使激活降低40％，而两个元件的突变均被取消。对于ACT缺失的突变体和CT突变为AG的突变体，也获得了相似的结果。 Vmw110对伊朗的激活也因AP-1元件的突变而减少（上游和下游元件分别为10倍和2倍），并且由于AP-1元件的突变而被取消。与AP-1顺式反应元件相邻的核苷酸突变对反式激活没有影响。用代表野生型ICP10启动子的DNA探针和HSV-1感染细胞的核提取物进行的凝胶阻滞分析鉴定出一种复合物，该复合物在模拟感染细胞或被Vmw110缺失突变体感染的细胞中看不到。当感染HSV-1的细胞与AP-1突变型DNA探针反应时，看不到该复合物，其形成与AP-1竞争，但与AP-1突变型寡核苷酸竞争。此复合物的迁移被c-Fos抗体阻滞，表明AP-1和Vmw110均参与其形成。 Vmw110也激活了在TATA框上游所有序列中缺失的突变体，但这种激活仅比野生型低2倍，而比双AP-1明显高（10倍）。突变体。数据表明，AP-1元素在ICP10基因表达/激活中起着至关重要的作用。

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