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Proceedings Paper

The HIV-1 Gag precursor induces the recruitment of Vpr oligomers to the plasma membrane as revealed by time-resolved fluorescence imaging
Author(s): Hugues de Rocquigny; Denis Dujardin; Tania Steffan; Pascal Didier; Yves Mély
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Paper Abstract

During formation of HIV particles, the Gag polyproteins are thought to interact with Vpr proteins to promote their encapsidation in the nascent particles. To directly visualize and monitor the formation of the Gag-Vpr complexes and correlate their formation with Vpr oligomerization, we used two photon lifetime imaging microscopy (FLIM) and time laps microscopy on HeLa cells expressing eGFP-labelled Vpr and tetracystein-tagged Gag proteins detected by the biarsenical ReAsH labelling reagent. Using these fluorescent microscopy approaches, we found that Gag proteins interact directly with Vpr proteins, which results in their transfer from the nuclear to the plasma membrane. Vpr oligomerization was found critical for both its interaction with Gag proteins and its transfer to the plasma membrane. Moreover, mutations in the C-terminal domain of Gag abolished the interaction with Vpr and its redistribution to the plasma membrane, indicating that this domain was critical for interaction with Vpr. Taken together, these data show that the Gag polyproteins interact through their C-terminal domain with Vpr oligomers, promoting the redistribution of the latter at the plasma membrane and probably their incorporation into nascent viral particles.

Paper Details

Date Published: 11 February 2011
PDF: 10 pages
Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032E (11 February 2011); doi: 10.1117/12.874052
Show Author Affiliations
Hugues de Rocquigny, Lab. de Biophotonique et Pharmacologie, CNRS, Univ. de Strasbourg (France)
Denis Dujardin, Lab. de Biophotonique et Pharmacologie, CNRS, Univ. de Strasbourg (France)
Tania Steffan, Lab. de Biophotonique et Pharmacologie, CNRS, Univ. de Strasbourg (France)
Pascal Didier, Lab. de Biophotonique et Pharmacologie, CNRS, Univ. de Strasbourg (France)
Yves Mély, Lab. de Biophotonique et Pharmacologie, CNRS, Univ. de Strasbourg (France)


Published in SPIE Proceedings Vol. 7903:
Multiphoton Microscopy in the Biomedical Sciences XI
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

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