Single molecule localisation microscopy reveals how HIV-1 Gag proteins sense membrane virus assembly sites in living host CD4 T cells
Sci Rep. 2018-11-02; 8(1):
DOI: 10.1038/s41598-018-34536-y
Lire sur PubMed
1. Sci Rep. 2018 Nov 2;8(1):16283. doi: 10.1038/s41598-018-34536-y.
Single molecule localisation microscopy reveals how HIV-1 Gag proteins sense
membrane virus assembly sites in living host CD4 T cells.
Floderer C(1), Masson JB(2), Boilley E(1), Georgeault S(3), Merida P(1), El
Beheiry M(4), Dahan M(4), Roingeard P(3), Sibarita JB(5), Favard C(6), Muriaux
D(7).
Author information:
(1)Infectious Disease Research Institute of Montpellier (IRIM), UMR9004 CNRS,
University of Montpellier, 1919 route de Mende, 34293, Montpellier, France.
(2)Decision and Bayesian Computation, UMR 3571 CNRS, Pasteur Institute, Paris,
France.
(3)INSERM U966 and IBiSA EM Facility, University of Tours, Tours, France.
(4)Light and Optical Control of Cellular Organization, Curie Institute, UMR, 168
CNRS, Paris, France.
(5)Interdisciplinary Institute for Neuroscience, UMR 5297 CNRS, University of
Bordeaux, Bordeaux, France.
(6)Infectious Disease Research Institute of Montpellier (IRIM), UMR9004 CNRS,
University of Montpellier, 1919 route de Mende, 34293, Montpellier, France.
.
(7)Infectious Disease Research Institute of Montpellier (IRIM), UMR9004 CNRS,
University of Montpellier, 1919 route de Mende, 34293, Montpellier, France.
.
Monitoring virus assembly at the nanoscale in host cells remains a major
challenge. Human immunodeficiency virus type 1 (HIV-1) components are addressed
to the plasma membrane where they assemble to form spherical particles of 100 nm
in diameter. Interestingly, HIV-1 Gag protein expression alone is sufficient to
produce virus-like particles (VLPs) that resemble the immature virus. Here, we
monitored VLP formation at the plasma membrane of host CD4+ T cells using a newly
developed workflow allowing the analysis of long duration recordings of
single-molecule Gag protein localisation and movement. Comparison of Gag
assembling platforms in CD4+ T cells expressing wild type or assembly-defective
Gag mutant proteins showed that VLP formation lasts roughly 15 minutes with an
assembly time of 5 minutes. Trapping energy maps, built from membrane associated
Gag protein movements, showed that one third of the assembling energy is due to
direct Gag capsid-capsid interaction while the remaining two thirds require the
nucleocapsid-RNA interactions. Finally, we show that the viral RNA genome does
not increase the attraction of Gag at the membrane towards the assembling site
but rather acts as a spatiotemporal coordinator of the membrane assembly process.
DOI: 10.1038/s41598-018-34536-y
PMCID: PMC6214999
PMID: 30389967