Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes

Nathalie Arhel (1) , Auguste Genovesio (2, 3) , Kim Ka (4) , Sarah Miko (1) , Emmanuelle Perret (5) , Jean-Christophe C Olivo-Marin (2) , Spencer L. Shorte (5) , Pierre Charneau (1)

1 Virologie moléculaire et Vectorologie
2 BIA - Analyse d'images biologiques - Biological Image Analysis
3 UPD5 - Université Paris Descartes - Paris 5
4 Institut Pasteur Korea - Institut Pasteur de Corée
5 PFID - Imagerie Dynamique (Plate-Forme)

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.

Domaines: Biochimie, Biologie Moléculaire

Nathalie Arhel, Auguste Genovesio, Kim Ka, Sarah Miko, Emmanuelle Perret, et al.. Quantitative four-dimensional tracking of cytoplasmic and nuclear HIV-1 complexes. Nature Methods, 2006, 3 (10), pp.817-824. ⟨10.1038/nmeth928⟩. ⟨pasteur-00163778⟩