Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes.
Emerging real-time techniques for imaging viral infections provide powerful tools for understanding the dynamics of virus-host cell interactions. Here we labeled human immunodeficiency virus-1 (HIV-1) integrase with a small tetracysteine tag, which preserved the virus' infectivity while allowing it to be labeled with the bis-arsenical fluorescein derivative FlAsH. This labeling allowed us to image both intracytoplasmic and intranuclear HIV-1 complexes in three dimensions over time (4D) in human cells and enabled us to analyze HIV-1 kinetics by automated 4D quantitative particle tracking. In the cytoplasm, HIV-1 complexes underwent directed movements toward the nuclear compartment, kinetically characteristic of both microtubule- and actin-dependent transport. The complexes then adopted smaller movements in a very confined volume once associated with the nuclear membrane and more diffuse movements once inside the nucleus. This work contributes new insight into the various movements of HIV-1 complexes within infected cells and provides a useful tool for the study of virus-host cell interactions during infection.
Arhel
Nathalie
N
Groupe de Virologie Moléculaire et Vectorologie, Institut Pasteur, 25-28 rue du Dr. Roux, 75724 Paris, France.
Genovesio
Auguste
A
Kim
Kyeong-Ae
KA
Miko
Sarah
S
Perret
Emmanuelle
E
Olivo-Marin
Jean-Christophe
JC
Shorte
Spencer
S
Charneau
Pierre
P
D001692:Biological Transport / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology / Q000821:virology; D002467:Cell Nucleus / Q000502:physiology / Q000821:virology*; D003545:Cysteine / Q000737:chemistry; D003593:Cytoplasm / Q000502:physiology / Q000821:virology*; D019427:HIV Integrase / Q000737:chemistry*; D015497:HIV-1 / Q000201:enzymology* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D012680:Sensitivity and Specificity; D013997:Time Factors
D019427:HIV Integrase; D003545:Cysteine