Characterization of the nuclear import pathway for HIV-1 integrase.
The karyophilic properties of the human immunodeficiency virus, type I (HIV-1) pre-integration complex (PIC) allow the virus to infect non-dividing cells. To better understand the mechanisms responsible for nuclear translocation of the PIC, we investigated nuclear import of HIV-1 integrase (IN), a PIC-associated viral enzyme involved in the integration of the viral genome in the host cell DNA. Accumulation of HIV-1 IN into nuclei of digitonin-permeabilized cells does not result from passive diffusion but rather from an active transport that occurs through the nuclear pore complexes. HIV-1 IN is imported by a saturable mechanism, implying that a limiting cellular factor is responsible for this process. Although IN has been previously proposed to contain classical basic nuclear localization signals, we found that nuclear accumulation of IN does not involve karyopherins alpha, beta1, and beta2-mediated pathways. Neither the non-hydrolyzable GTP analog, guanosine 5'-O-(thiotriphosphate), nor the GTP hydrolysis-deficient Ran mutant, RanQ69L, significantly affects nuclear import of IN, which depends instead on ATP hydrolysis. Therefore these results support the idea that IN import is not mediated by members of the karyopherin beta family. More generally, in vitro nuclear import of IN does not require addition of cytosolic factors, suggesting that cellular factor(s) involved in this active but atypical pathway process probably remain associated with the nuclear compartment or the nuclear pore complexes from permeabilized cells.
Institut Jacques Monod, Unité Mixte de Recherche 7592, CNRS, Université Paris VI, Université Paris VII, Paris 75251, France.
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Depienne
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Mousnier
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Leh
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Le Rouzic
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Dormont
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Benichou
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Dargemont
D001692:Biological Transport; D002467:Cell Nucleus / Q000378:metabolism*; D015658:HIV Infections / Q000821:virology; D019427:HIV Integrase / Q000378:metabolism*; D015497:HIV-1 / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D014779:Virus Replication
D019427:HIV Integrase