The tyrosine kinase Hck is an inhibitor of HIV-1 replication counteracted by the viral vif protein.
The virus infectivity factor (Vif) protein facilitates the replication of human immunodeficiency virus type 1 (HIV-1) in primary lymphocytes and macrophages. Its action is strongly dependent on the cellular environment, and it has been proposed that the Vif protein counteracts cellular activities that would otherwise limit HIV-1 replication. Using a glutathione S-transferase pull-down assay, we identified that Vif binds specifically to the Src homology 3 domain of Hck, a tyrosine kinase from the Src family. The interaction between Vif and the full-length Hck was further assessed by co-precipitation assays in vitro and in human cells. The Vif protein repressed the kinase activity of Hck and was not itself a substrate for Hck phosphorylation. Within one single replication cycle of HIV-1, Hck was able to inhibit the production and the infectivity of vif-deleted virus but not that of wild-type virus. Accordingly, HIV-1 vif- replication was delayed in Jurkat T cell clones stably expressing Hck. Our data demonstrate that Hck controls negatively HIV-1 replication and that this inhibition is suppressed by the expression of Vif. Hck, which is present in monocyte-macrophage cells, represents the first identified cellular inhibitor of HIV-1 replication overcome by Vif.
INSERM Unit 372, Université de la Méditerranée, 163 Avenue de Luminy, 13276 Marseilles Cedex 09, France.
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Hassaïne
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Courcoul
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Bessou
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Barthalay
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Picard
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Olive
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Collette
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Vigne
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Decroly
D000595:Amino Acid Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D016384:Consensus Sequence; D016346:Gene Products, vif / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016341:Genes, vif; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D007668:Kidney; D007700:Kinetics; D008969:Molecular Sequence Data; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051572:Proto-Oncogene Proteins c-hck; D011519:Proto-Oncogenes; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D020298:U937 Cells; D014779:Virus Replication / Q000502:physiology*; D018909:src Homology Domains; D054320:vif Gene Products, Human Immunodeficiency Virus
D016346:Gene Products, vif; D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; D054320:vif Gene Products, Human Immunodeficiency Virus; D011505:Protein-Tyrosine Kinases; C496631:HCK protein, human; D051572:Proto-Oncogene Proteins c-hck