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The intracellular plasma membrane-connected compartment in the assembly of HIV-1 in human macrophages

Nkwe, DO; Pelchen-Matthews, A; Burden, JJ; Collinson, LM; Marsh, M; (2016) The intracellular plasma membrane-connected compartment in the assembly of HIV-1 in human macrophages. BMC Biology , 14 (50) 10.1186/s12915-016-0272-3. Green open access

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Abstract

Background In HIV-infected macrophages, newly formed progeny virus particles accumulate in intracellular plasma membrane-connected compartments (IPMCs). Although the virus is usually seen in these compartments, it is unclear whether HIV assembly is specifically targeted to IPMCs or whether some viruses may also form at the cell surface but are not detected, as particles budding from the latter site will be released into the medium. Results To investigate the fidelity of HIV-1 targeting to IPMCs compared to the cell surface directly, we generated mutants defective in recruitment of the Endosomal Sorting Complexes Required for Transport (ESCRT) proteins required for virus scission. For mutants unable to bind the ESCRT-I component Tsg101, HIV release was inhibited and light and electron microscopy revealed that budding was arrested. When expressed in human monocyte-derived macrophages (MDM), these mutants formed budding-arrested, immature particles at their assembly sites, allowing us to capture virtually all of the virus budding events. A detailed morphological analysis of the distribution of the arrested viruses by immunofluorescence staining and confocal microscopy, and by electron microscopy, demonstrated that HIV assembly in MDMs is targeted primarily to IPMCs, with fewer than 5 % of budding events seen at the cell surface. Morphometric analysis of the relative membrane areas at the cell surface and IPMCs confirmed a large enrichment of virus assembly events in IPMCs. Serial block-face scanning electron microscopy of macrophages infected with a budding-defective HIV mutant revealed high-resolution 3D views of the complex organisation of IPMCs, with in excess of 15,000 associated HIV budding sites, and multiple connections between IPMCs and the cell surface.

Type: Article
Title: The intracellular plasma membrane-connected compartment in the assembly of HIV-1 in human macrophages
Open access status: An open access version is available from UCL Discovery
DOI: 10.1186/s12915-016-0272-3
Publisher version: http://dx.doi.org/10.1186/s12915-016-0272-3
Additional information: This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Keywords: Science & Technology, Life Sciences & Biomedicine, Biology, Life Sciences & Biomedicine - Other Topics, HIV-1, Assembly compartment, Macrophage, Intracellular plasma membrane-connected compartments, Endosomal sorting complexes required for transport, Volume electron microscopy, T-CELLS, NEUROCOGNITIVE DISORDERS, NEUTRALIZING ANTIBODIES, GAG, INFECTION, PROTEIN, DOMAIN, NEUROPATHOGENESIS, TRANSMISSION, MECHANISMS
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Pop Health Sciences > Institute for Global Health
URI: https://discovery.ucl.ac.uk/id/eprint/1506007
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