Repression of the human immunodeficiency virus type 1 promoter by the human KRAB domain results in inhibition of virus production
Repression of the human immunodeficiency virus type 1 promoter by the human KRAB domain results in inhibition of virus production
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DOI：
10.1016/s0167-4781(99)00046-9
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发表时间：
1999-05-14
期刊：
影响因子：
--
通讯作者：
Schneider, J
中科院分区：
文献类型：
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作者：
Herchenröder, O;Hahne, JC;Schneider, J
Kruppel 相关盒 (KRAB) 结构域被描述为真核转录抑制因子。我们表明，KRAB 与 DNA 结合域的融合提供了一种抑制具有复制能力的人类免疫缺陷病毒 (HN) 基因组表达的新方法。人锌指蛋白 KOX1 的 KRAB 结构域与大肠杆菌四环素阻遏物 (TetR) 的 DNA 结合结构域相结合。 HeLa 细胞中 TetR-KRAB 蛋白的组成型表达可抑制编码 TetR 靶序列的 HIV 基因组产生病毒 80%。 HIV启动子驱动的报告质粒也观察到了相同的抑制作用。通过提供信息的 KRAB 突变体、缺乏相应靶序列的质粒以及通过用四环素逆转 TetR-KRAB 介导的抑制来显示抑制的特异性。 TetR-KRAB 在 6 kbp 处与启动子结合，从而抑制病毒产生。因此，我们得出结论，真正的 HIV 基因组的任何位点都可以作为嵌合 KRAB 阻遏蛋白的靶标。通过人工选择的结合结构域对 KRAB 结构域的特异性靶向可能通常适用于控制哺乳动物细胞中的转录。 (C) 1999 Elsevier Science B.V. 保留所有权利。
The Kruppel-associated box (KRAB) domain has been described as a eukaryotic repressor of transcription. We show that fusion of KRAB to DNA-binding-domains provides a novel approach to inhibit expression of a replication-competent human immunodeficiency virus (HN) genome. The KRAB domain from the human zinc finger protein KOX1 was combined with the DNA binding domain of the Escherichia coli tetracycline repressor (TetR). Constitutive expression of the TetR-KRAB protein in HeLa cells inhibited virus production from an HIV genome encoding TetR target sequences by 80%. The same inhibition was observed with HIV-promoter-driven reporter plasmids. The specificity of inhibition was shown with informative KRAB mutants, plasmids lacking the respective target sequences, and by reversal of the TetR-KRAB-mediated inhibition with tetracycline. Virus production was suppressed by binding of TetR-KRAB at a distance of 6 kbp to the promoter. We therefore conclude that any site of the genuine HIV genome could serve as target of a chimeric KRAB repressor protein. Specific targeting of the KRAB domain by artificially selected binding domains may be generally applicable to control transcription in mammalian cells. (C) 1999 Elsevier Science B.V. All rights reserved.