Nucleocapsid Promotes Localization of HIV-1 Gag to Uropods That Participate in Virological Synapses between T Cells

PLoS Pathogens

Akira Ono¹  G. Nicholas Llewellyn¹  Jonathan R. Grover²  Ian B. Hogue² 

[1] Cellular and Molecular Biology Program, University of Michigan Medical School, Ann Arbor, Michigan, United States of America;Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, United States of America

关键词: T cells;    HIV-1;    Cell membranes;    Fluorescence resonance energy transfer;    Dimerization;    Membrane proteins;    Polarized light microscopy;    Synapses;   

DOI  :  10.1371/journal.ppat.1001167

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

T cells adopt a polarized morphology in lymphoid organs, where cell-to-cell transmission of HIV-1 is likely frequent. However, despite the importance of understanding virus spread in vivo, little is known about the HIV-1 life cycle, particularly its late phase, in polarized T cells. Polarized T cells form two ends, the leading edge at the front and a protrusion called a uropod at the rear. Using multiple uropod markers, we observed that HIV-1 Gag localizes to the uropod in polarized T cells. Infected T cells formed contacts with uninfected target T cells preferentially via HIV-1 Gag-containing uropods compared to leading edges that lack plasma-membrane-associated Gag. Cell contacts enriched in Gag and CD4, which define the virological synapse (VS), are also enriched in uropod markers. These results indicate that Gag-laden uropods participate in the formation and/or structure of the VS, which likely plays a key role in cell-to-cell transmission of HIV-1. Consistent with this notion, a myosin light chain kinase inhibitor, which disrupts uropods, reduced virus particle transfer from infected T cells to target T cells. Mechanistically, we observed that Gag copatches with antibody-crosslinked uropod markers even in non-polarized cells, suggesting an association of Gag with uropod-specific microdomains that carry Gag to uropods. Finally, we determined that localization of Gag to the uropod depends on higher-order clustering driven by its NC domain. Taken together, these results support a model in which NC-dependent Gag accumulation to uropods establishes a preformed platform that later constitutes T-cell-T-cell contacts at which HIV-1 virus transfer occurs.

【 授权许可】

CC BY