Nuclear localization of human immunodeficiency virus type 1 integrase expressed as a fusion protein with green fluorescent protein.
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.
Pluymers
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Rega Institute for Medical Research, Katholieke Universiteit, Leuven, Belgium.
Cherepanov
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P
Schols
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D
De Clercq
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E
Debyser
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Z
D002455:Cell Division; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology* / Q000821:virology; D005434:Flow Cytometry; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D019427:HIV Integrase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology*; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D019427:HIV Integrase