The HIV-1 Gag precursor induces the recruitment of Vpr oligomers to the plasma membrane as revealed by time-resolved fluorescence imaging

Hugues de Rocquigny; Denis Dujardin; Tania Steffan; Pascal Didier; Yves Mély

doi:10.1117/12.874052

11 February 2011 The HIV-1 Gag precursor induces the recruitment of Vpr oligomers to the plasma membrane as revealed by time-resolved fluorescence imaging

Hugues de Rocquigny, Denis Dujardin, Tania Steffan, Pascal Didier, Yves Mély

Author Affiliations +

Proceedings Volume 7903, Multiphoton Microscopy in the Biomedical Sciences XI; 79032E (2011) https://doi.org/10.1117/12.874052
Event: SPIE BiOS, 2011, San Francisco, California, United States

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.

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Hugues de Rocquigny, Denis Dujardin, Tania Steffan, Pascal Didier, and Yves Mély "The HIV-1 Gag precursor induces the recruitment of Vpr oligomers to the plasma membrane as revealed by time-resolved fluorescence imaging", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032E (11 February 2011); https://doi.org/10.1117/12.874052

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KEYWORDS

Proteins

Luminescence

Fluorescence lifetime imaging

Plasma

Microscopy

Particles

Fluorescence resonance energy transfer

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