title: Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes.
abstract: 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.
authors.affiliation: Groupe de Virologie Moléculaire et Vectorologie, Institut Pasteur, 25-28 rue du Dr. Roux, 75724 Paris, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Arhel
authors.forename: Auguste
authors.initials: A
authors.lastname: Genovesio
authors.forename: Kyeong-Ae
authors.initials: KA
authors.lastname: Kim
authors.forename: Sarah
authors.initials: S
authors.lastname: Miko
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Perret
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Olivo-Marin
authors.forename: Spencer
authors.initials: S
authors.lastname: Shorte
authors.forename: Pierre
authors.initials: P
authors.lastname: Charneau
mesh_terms: 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
chemical_list: D019427:HIV Integrase; D003545:Cysteine