Pluymers W, Cherepanov P, Schols D, De Clercq E, Debyser Z
Rega Institute for Medical Research, Katholieke Universiteit, Leuven, Belgium.
Virology. 1999 Jun 5;258(2):327-32. doi: 10.1006/viro.1999.9727.
Lentiviruses in general and the human immunodeficiency virus type 1 (HIV-1) in particular have the ability to integrate their genome stably into the chromosome of nondividing cells. Integration of HIV cDNA is mediated by the viral integrase (IN). Apart from its catalytic activity, this enzyme seems to play an important role in the transport of the HIV preintegration complex into the nucleus of nondividing cells. We studied the karyophilic properties of IN by constructing an N-terminal fusion protein of HIV-1 integrase and green fluorescent protein (GFP-IN). Transient expression of GFP-IN in various mammalian cell lines was demonstrated by fluorescence microscopy, flow cytometry, and Western blotting. Although wild-type GFP was localized throughout the cell, GFP-IN was localized predominantly in the nucleus. Nuclear localization of GFP-IN was also obtained after transient transfection of the cells arrested in the G1/S phase of the cell cycle. These results provide compelling evidence for the karyophilic properties of the HIV-1 integrase.
一般来说，慢病毒，尤其是人类免疫缺陷病毒1型（HIV-1），能够将其基因组稳定整合到非分裂细胞的染色体中。HIV cDNA的整合由病毒整合酶（IN）介导。除了其催化活性外，这种酶似乎在将HIV前整合复合物转运到非分裂细胞的细胞核中起着重要作用。我们通过构建HIV-1整合酶与绿色荧光蛋白的N端融合蛋白（GFP-IN）来研究整合酶的亲核特性。通过荧光显微镜、流式细胞术和蛋白质印迹法证实了GFP-IN在各种哺乳动物细胞系中的瞬时表达。尽管野生型GFP分布于整个细胞，但GFP-IN主要定位于细胞核。在细胞周期的G1/S期停滞的细胞中进行瞬时转染后，也获得了GFP-IN的核定位。这些结果为HIV-1整合酶的亲核特性提供了令人信服的证据。
