Nature Methods
- 3, 817 - 824 (2006)
Published online: 21 September 2006; | doi:10.1038/nmeth928
Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexesNathalie Arhel1, 6, Auguste Genovesio2, 4, 6, Kyeong-Ae Kim5, Sarah Miko1, Emmanuelle Perret3, Jean-Christophe Olivo-Marin2, Spencer Shorte3 & Pierre Charneau11
Groupe de Virologie Moléculaire et Vectorologie. 2
Unité d'Analyse d'Images Quantitative. 3
Centre d'Imagerie Dynamique, Institut Pasteur, 25-28 rue du Dr. Roux, 75724 Paris, France. 4
SIP-CRIP5 Université Paris V, 45 rue des Saints Pères, 75006 Paris, France. 5
Institut Pasteur Korea, 39-1, Hawolgok-dong, Sungbuk-gu, Seoul, Korea. 6
These authors contributed equally to this work.
Correspondence should be addressed to Pierre Charneau charneau@pasteur.fr or Jean-Christophe Olivo-Marin jcolivo@pasteur.fr 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.
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