Real-Time Visualization of HIV-1 GAG Trafficking in Infected Macrophages

PLoS Pathogens

David E. Ott¹  Lori V. Coren¹  Akira Ono²  Ferri Soheilian³  Kunio Nagashima³  Eric O. Freed⁴  Sherimay D. Ablan⁴  Karine Gousset⁴ 

[1] AIDS Vaccine Program, SAIC-Frederick, Inc., National Cancer Institute, Frederick, Maryland, United States of America;Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, United States of America;Image Analysis Laboratory, Advanced Technology Program, SAIC-Frederick, National Cancer Institute at Frederick, Frederick, Maryland, United States of America;Virus-Cell Interaction Section, HIV Drug Resistance Program, National Cancer Institute, Frederick, Maryland, United States of America

关键词: Synapses;    T cells;    HIV-1;    Macrophages;    Virions;    HeLa cells;    Viral replication;    Cell staining;   

DOI  :  10.1371/journal.ppat.1000015

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

HIV-1 particle production is driven by the Gag precursor protein Pr55Gag. Despite significant progress in defining both the viral and cellular determinants of HIV-1 assembly and release, the trafficking pathway used by Gag to reach its site of assembly in the infected cell remains to be elucidated. The Gag trafficking itinerary in primary monocyte-derived macrophages is especially poorly understood. To define the site of assembly and characterize the Gag trafficking pathway in this physiologically relevant cell type, we have made use of the biarsenical-tetracysteine system. A small tetracysteine tag was introduced near the C-terminus of the matrix domain of Gag. The insertion of the tag at this position did not interfere with Gag trafficking, virus assembly or release, particle infectivity, or the kinetics of virus replication. By using this in vivo detection system to visualize Gag trafficking in living macrophages, Gag was observed to accumulate both at the plasma membrane and in an apparently internal compartment that bears markers characteristic of late endosomes or multivesicular bodies. Significantly, the internal Gag rapidly translocated to the junction between the infected macrophages and uninfected T cells following macrophage/T-cell synapse formation. These data indicate that a population of Gag in infected macrophages remains sequestered internally and is presented to uninfected target cells at a virological synapse.

【 授权许可】

CC BY