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TitleDirect visualization of HIV-1 core nuclear import and its interplay with the nuclear pore.
Journal, issue, pagesEMBO Rep, Vol. 26, Issue 21, Page 5133-5153, Year 2025
Publish dateAug 29, 2025
AuthorsZhen Hou / Stanley Fronik / Yao Shen / Long Chen / Christopher Thompson / Sarah Neumann / Peijun Zhang /
PubMed AbstractDirect 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 ...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.
External linksEMBO Rep / PubMed:40883508 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution12.5 - 38.0 Å
Structure data

EMDB-53458: In situ cryo-EM structure of HIV-1 VLP CA hexamer before the nuclear import
Method: EM (subtomogram averaging) / Resolution: 12.5 Å

EMDB-53459: In situ cryo-EM structure of HIV-1 VLP hexamer in the NPC
Method: EM (subtomogram averaging) / Resolution: 13.5 Å

EMDB-53460: In situ cryo-EM structure of HIV-1 VLP hexamer after the nuclear import
Method: EM (subtomogram averaging) / Resolution: 13.5 Å

EMDB-54198: In-situ structure of cytoplasmic ring of NPC of CEM T lymphoblast cell
Method: EM (subtomogram averaging) / Resolution: 32.0 Å

EMDB-54199: In-situ structure of inner ring of NPC of CEM T lymphoblast
Method: EM (subtomogram averaging) / Resolution: 38.0 Å

Source
  • Homo sapiens (human)

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