Cytoskeleton association and virion incorporation of the human immunodeficiency virus type 1 Vif protein.
The human immunodeficiency virus type 1 (HIV-1) Vif protein has an important role in the regulation of virus infectivity. This function of Vif is cell type specific, and virions produced in the absence of Vif in restrictive cells have greatly reduced infectivity. We show here that the intracellular localization of Vif is dependent on the presence of the intermediate filament vimentin. Fractionation of acutely infected T cells or transiently transfected HeLa cells demonstrates the existence of a soluble and a cytoskeletal form and to a lesser extent the presence of a detergent-extractable form of Vif. Confocal microscopy suggests that in HeLa cells, Vif is predominantly present in the cytoplasm and closely colocalizes with the intermediate filament vimentin. Treatment of cells with drugs affecting the structure of vimentin filaments affect the localization of Vif accordingly, indicating a close association of Vif with this cytoskeletal component. The association of Vif with vimentin can cause the collapse of the intermediate filament network into a perinuclear aggregate. In contrast, analysis of Vif in vimentin-negative cells reveals significant staining of the nucleus and the nuclear membrane in addition to diffuse cytoplasmic staining. In addition to the association of Vif with intermediate filaments, analyses of virion preparations demonstrate that Vif is incorporated into virus particles. In sucrose density gradients, Vif cosediments with capsid proteins even after detergent treatment of virus preparations, suggesting that Vif is associated with the inner core of HIV particles. We propose a model in which Vif has a crucial function as a virion component either by regulating virus maturation or following virus entry into a host cell possibly involving an interaction with the cellular cytoskeletal network.
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892-0460, USA.
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Karczewski
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Strebel
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism / Q000821:virology*; D004279:DNA, Viral; D016346:Gene Products, vif / Q000378:metabolism*; D006679:HIV Seropositivity / Q000097:blood / Q000276:immunology; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000378:metabolism*; D014771:Virion / Q000378:metabolism; D054320:vif Gene Products, Human Immunodeficiency Virus
D004279:DNA, Viral; D016346:Gene Products, vif; D014746:Vimentin; D054320:vif Gene Products, Human Immunodeficiency Virus