Year: 2025
Type: article
Abstract: Direct visualization of HIV-1 nuclear import through the nuclear pore complex (NPC) presents a technical challenge due to the rarity of this process. To enable systematic investigation, we developed a robust in situ system that mimics HIV-1 nuclear import in a near-native context using isolated HIV-1 virus like particles (VLP) cores and permeabilized CD4 + T lymphocyte (CEM) cells. This approach supports docking and translocation of abundant viral cores through nuclear pores into the nucleus. For high-resolution visualization, we implemented an integrated correlative approach to guide cryo-focused ion beam (cryo-FIB) milling and cryo-electron tomography (cryo-ET) imaging, enabling precise targeting and structural characterization of individual nuclear import events. Using this workflow, we visualized 510 HIV-1 VLP cores at distinct stages of nuclear import, capturing key snapshots of the full progression of nuclear import. Subsequent statistical and structural analyses allow classification of core morphologies and identification of translocation-associated remodeling in nuclear pores. This work provides a methodological foundation for dissecting HIV-1 and potentially other viruses nuclear import processes and post-entry events in a controlled and quantitative manner.
Source: EMBO Reports
Authors: Zhen Hou, Stanley Fronik, Yao Shen, Long Chen, Christopher Thompson, Sarah Neumann, Peijun Zhang
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $7,990
Awards: Human Genetics and Disease Biology: Core Renewal for the Wellcome Trust Centre for Human Genetics, G3400220936, Molecular mechanisms of HIV-1 restriction by capsid-sensing host cell proteins, G3954569211, Assembly and Dynamics of Bacterial Chemosensory Signaling Arrays, Resolving HIV-1 transport and host cofactor regulation in the cellular context, Molecular mechanisms of HIV-1 restriction by capsid-sensing host cell proteins, G8364292457, G839554599, Pittsburgh Center for HIV Protein Interactions (PCHPI)
OpenAlex ID: W4413833087