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Endosome-to-cytosol transport of viral nucleocapsids

Abstract

During viral infection, fusion of the viral envelope with endosomal membranes and nucleocapsid release were thought to be concomitant events. We show here that for the vesicular stomatitis virus they occur sequentially, at two successive steps of the endocytic pathway. Fusion already occurs in transport intermediates between early and late endosomes, presumably releasing the nucleocapsid within the lumen of intra-endosomal vesicles, where it remains hidden. Transport to late endosomes is then required for the nucleocapsid to be delivered to the cytoplasm. This last step, which initiates infection, depends on the late endosomal lipid lysobisphosphatidic acid (LBPA) and its putative effector Alix/AIP1, and is regulated by phosphatidylinositol-3-phosphate (PtdIns(3)P) signalling via the PtdIns(3)P-binding protein Snx16. We conclude that the nucleocapsid is exported into the cytoplasm after the back-fusion of internal vesicles with the limiting membrane of late endosomes, and that this process is controlled by the phospholipids LBPA and PtdIns(3)P and their effectors.

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Figure 1: Fusion of individual viral particles in the endocytic pathway.
Figure 2: Role of microtubules in viral fusion and nucleocapsid release.
Figure 3: VSV infection requires transport to late endosomes.
Figure 4: Treatments that affect microtubules and LBPA reduce nucelocapsid release and viral protein synthesis.
Figure 5: Viral fusion and nucleocapsid release after PI(3)K inhibition.
Figure 6: Viral nucleocapsids reside within intraluminal endosomal vesicles.
Figure 7: Role of Hrs and PtdIns(3)P in viral fusion and nucleocapsid release.
Figure 8: Reconstitution of nucleocapsid release using an in vitro assay.

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Acknowledgements

We would like to thank B. Viaccoz, M.-C. Velluz and M.-H. Beuchat for expert technical assistance, and U. Laemmli and C. Bauer (Bioimaging platform, Frontiers in Genetics NCCR, Geneva) for help with light microscopy and imaging. We are grateful to K. Miwa and D. Trono for providing us with concanamycin B and HIV-1-derived vector pseudotyped with VSV-G, respectively. We also wish to thank G. van der Goot and M. Fivaz for critical reading of the manuscript. This work was supported by the Swiss National Science Foundation (J.G.), the International Human Frontier Science Program (J.G and R.G.P.), and the Australian National Health and Medical Research Council (R.G.P.). I.L.B. was supported by the award of the Bettencourt Foundation price (Prix des Jeunes Chercheurs), by the French Cancer Research Association (ARC) and by the Roche Foundation. V.P. was supported by the Roche Foundation and EMBO.

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Le Blanc, I., Luyet, PP., Pons, V. et al. Endosome-to-cytosol transport of viral nucleocapsids. Nat Cell Biol 7, 653–664 (2005). https://doi.org/10.1038/ncb1269

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