Visualization of the intracellular behavior of HIV in living cells.
To track the behavior of human immunodeficiency virus (HIV)-1 in the cytoplasm of infected cells, we have tagged virions by incorporation of HIV Vpr fused to the GFP. Observation of the GFP-labeled particles in living cells revealed that they moved in curvilinear paths in the cytoplasm and accumulated in the perinuclear region, often near the microtubule-organizing center. Further studies show that HIV uses cytoplasmic dynein and the microtubule network to migrate toward the nucleus. By combining GFP fused to the NH2 terminus of HIV-1 Vpr tagging with other labeling techniques, it was possible to determine the state of progression of individual particles through the viral life cycle. Correlation of immunofluorescent and electron micrographs allowed high resolution imaging of microtubule-associated structures that are proposed to be reverse transcription complexes. Based on these observations, we propose that HIV uses dynein and the microtubule network to facilitate the delivery of the viral genome to the nucleus of the cell during early postentry steps of the HIV life cycle.
McDonald
David
D
Dept. of Microbiology and Immunology, University of Illinois, 835 South Wolcott Avenue, Chicago, IL 60612, USA.
Vodicka
Marie A
MA
Lucero
Ginger
G
Svitkina
Tatyana M
TM
Borisy
Gary G
GG
Emerman
Michael
M
Hope
Thomas J
TJ
D001692:Biological Transport / Q000502:physiology*; D015704:CD4 Antigens / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology*; D003593:Cytoplasm / Q000378:metabolism / Q000821:virology*; D003599:Cytoskeleton / Q000378:metabolism / Q000821:virology; D004398:Dyneins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D015683:Gene Products, gag / Q000378:metabolism; D016336:Gene Products, vpr / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D016655:HIV Core Protein p24 / Q000378:metabolism; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D046911:Macromolecular Substances; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008870:Microtubules / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014158:Transcription, Genetic; D014771:Virion / Q000235:genetics / Q000502:physiology*; D054325:vpr Gene Products, Human Immunodeficiency Virus
D015704:CD4 Antigens; D005456:Fluorescent Dyes; D015683:Gene Products, gag; D016336:Gene Products, vpr; D016655:HIV Core Protein p24; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D046911:Macromolecular Substances; D020409:Molecular Motor Proteins; D011993:Recombinant Fusion Proteins; D054325:vpr Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins; D004398:Dyneins