TETHERING HUMAN-IMMUNODEFICIENCY-VIRUS-1 INTEGRASE TO A DNA SITE DIRECTS INTEGRATION TO NEARBY SEQUENCES
TETHERING HUMAN-IMMUNODEFICIENCY-VIRUS-1 INTEGRASE TO A DNA SITE DIRECTS INTEGRATION TO NEARBY SEQUENCES
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DOI：
10.1073/pnas.91.20.9233
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发表时间：
1994-09-27
影响因子：
11.1
通讯作者：
BUSHMAN, FD
中科院分区：
文献类型：
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作者：
BUSHMAN, FD
某些逆转录病毒和逆转录转座子在选择用于整合的宿主DNA位点方面显示出强烈的偏好。为了探索逆转录元件整合酶蛋白与靶DNA位点的简单拴系足以指导整合的可能性，分析了由人免疫缺陷病毒1整合酶和λ阻遏物组成的杂合物的活性。在含有几个目标DNA的体外反应中，发现λ阻遏物-整合酶杂合体选择性地直接整合到含有λ操纵子的目标。添加λ阻遏物阻断了选择性整合，表明需要与操作员结合。λ阻遏物-整合酶杂合蛋白主要将整合定向到B-DNA螺旋的同一面上的操纵子附近的位点，这表明靶DNA可能是通过环出间插序列而被捕获的。这种杂合整合酶蛋白可用于在体内将逆转录病毒整合到特定序列。
Certain retrovirus and retrotransposons display strong biases in the selection of host DNA sites for integration. To probe the possibility that simple tethering of the retroelement integrase protein to a target DNA site is sufficient to direct integration, the activities of a hybrid composed of human immunodeficiency virus 1 integrase and lambda repressor were analyzed. In in vitro reactions containing several target DNAs, the lambda repressor-integrase hybrid was found to direct integration selectively to targets containing lambda operators. Addition of lambda repressor blocked selective integration, indicating that binding to the operators was required. The lambda repressor-integrase hybrid protein directed integration primarily to sites near the operators on the same face of the B-DNA helix, indicating that target DNA was probably captured by looping out the intervening sequences. Such hybrid integrase proteins may be useful for directing retroviral integration to specific sequences in vivo.