Vink C, Oude Groeneger A M, Plasterk R H
Division of Molecular Biology, The Netherlands Cancer Institute, Amsterdam.
Nucleic Acids Res. 1993 Mar 25;21(6):1419-25. doi: 10.1093/nar/21.6.1419.
The integrase (IN) protein of the human immunodeficiency virus (HIV) is required for specific cleavage of the viral DNA termini, and subsequent integration of the viral DNA into target DNA. To identify the various domains of the IN protein we generated a series of IN deletion mutants as fusions to maltose-binding protein (MBP). The deletion mutants were tested for their ability to bind DNA, to mediate site-specific cleavage of the viral DNA ends, and to carry out integration and disintegration reactions. We found that the DNA-binding region resides between amino acids 200 and 270 of the 288-residues HIV-1 IN protein. The catalytic domain of the protein was mapped between amino acids 50 and 194. For the specific activities of IN, cleavage of the viral DNA and integration, both the DNA-binding domain and the conserved amino-terminal region of IN are required. These regions are dispensable however, for disintegration activity.
人类免疫缺陷病毒（HIV）的整合酶（IN）蛋白对于病毒DNA末端的特异性切割以及随后病毒DNA整合到靶DNA中是必需的。为了鉴定IN蛋白的各个结构域，我们构建了一系列与麦芽糖结合蛋白（MBP）融合的IN缺失突变体。测试这些缺失突变体结合DNA的能力、介导病毒DNA末端位点特异性切割的能力以及进行整合和解离反应的能力。我们发现DNA结合区域位于288个氨基酸残基的HIV-1 IN蛋白的第200至270个氨基酸之间。该蛋白的催化结构域定位在第50至194个氨基酸之间。对于IN的特异性活性，即病毒DNA的切割和整合，IN的DNA结合结构域和保守的氨基末端区域都是必需的。然而，这些区域对于解离活性是可有可无的。
