Intracellular transport of human immunodeficiency virus type 1 genomic RNA and viral production are dependent on dynein motor function and late endosome positioning.
Our earlier work indicated that the human immunodeficiency virus type 1 (HIV-1) genomic RNA (vRNA) is trafficked to the microtubule-organizing center (MTOC) when heterogeneous nuclear ribonucleoprotein A2/B1 is depleted from cells. Also, Rab7-interacting lysosomal protein promoted dynein motor complex, late endosome and vRNA clustering at the MTOC suggesting that the dynein motor and late endosomes were involved in vRNA trafficking. To investigate the role of the dynein motor in vRNA trafficking, dynein motor function was disrupted by small interference RNA-mediated depletion of the dynein heavy chain or by p50/dynamitin overexpression. These treatments led to a marked relocalization of vRNA and viral structural protein Gag to the cell periphery with late endosomes and a severalfold increase in HIV-1 production. In contrast, rerouting vRNA to the MTOC reduced virus production. vRNA localization depended on Gag membrane association as shown using both myristoylation and Gag nucleocapsid domain proviral mutants. Furthermore, the cytoplasmic localization of vRNA and Gag was not attributable to intracellular or internalized endocytosed virus particles. Our results demonstrate that dynein motor function is important for regulating Gag and vRNA egress on endosomal membranes in the cytoplasm to directly impact on viral production.
Lehmann
Martin
M
HIV-1 RNA Trafficking Laboratory, Lady Davis Institute for Medical Research, Quebec.
Milev
Miroslav P
MP
Abrahamyan
Levon
L
Yao
Xiao-Jian
XJ
Pante
Nelly
N
Mouland
Andrew J
AJ
D000595:Amino Acid Sequence; D001692:Biological Transport; D004398:Dyneins / Q000378:metabolism*; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology*; D015683:Gene Products, gag / Q000737:chemistry / Q000378:metabolism; D016679:Genome, Viral / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics* / Q000648:ultrastructure; D006367:HeLa Cells; D054298:Human Immunodeficiency Virus Proteins / Q000378:metabolism; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D042541:Intracellular Space / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology*; D052118:Lysosomal-Associated Membrane Protein 1 / Q000378:metabolism; D008969:Molecular Sequence Data; D019814:Myristic Acid / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D054334:Viral Regulatory and Accessory Proteins / Q000378:metabolism; D014771:Virion / Q000378:metabolism; D019065:Virus Assembly; D000086082:Viroporin Proteins
D004398:Dyneins; D015683:Gene Products, gag; D054298:Human Immunodeficiency Virus Proteins; D052118:Lysosomal-Associated Membrane Protein 1; D019814:Myristic Acid; D012367:RNA, Viral; D054334:Viral Regulatory and Accessory Proteins; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins