HIV-induced membraneless organelles orchestrate post-nuclear entry steps

Journal of Molecular Cell Biology, Год журнала: 2022, Номер 14(11)

Опубликована: Окт. 31, 2022

ABSTRACT HIV integration occurs in chromatin sites that favor the release of high levels viral progeny; alternatively, virus is also able to discreetly coexist with host. The infection perturbs cellular environment inducing remodelling nuclear landscape. Indeed, HIV-1 triggers clustering host factor CPSF6, but underlying mechanism poorly understood. Our data indicate usurps a recently discovered biological phenomenon, called liquid–liquid phase separation, hijack cell. We observed CPSF6 clusters as part HIV-induced membraneless organelles (HIV-1 MLOs) macrophages, one main target cell types. describe MLOs follow phase-separation rules and represent functional biomolecular condensates. highlight hubs reverse transcription, while double-stranded DNA, once formed, rapidly migrates outside these structures. Transcription-competent proviruses localize near LEDGF-abundant regions, known be active sites. Therefore, orchestrate events prior step create favorable for replication. This study uncovers single host–viral complexes their landscape, which markedly restructured by HIV-1.

Язык: Английский

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IP6‐stabilised HIV capsids evade cGAS/STING‐mediated host immune sensing

EMBO Reports, Год журнала: 2023, Номер 24(5)

Опубликована: Март 27, 2023

Abstract HIV‐1 uses inositol hexakisphosphate (IP6) to build a metastable capsid capable of delivering its genome into the host nucleus. Here, we show that viruses are unable package IP6 lack protection and detected by innate immunity, resulting in activation an antiviral state inhibits infection. Disrupting enrichment results defective capsids trigger cytokine chemokine responses during infection both primary macrophages T‐cell lines. Restoring with single mutation rescues ability infect cells without being detected. Using combination mutants CRISPR‐derived knockout cell lines for RNA DNA sensors, immune sensing is dependent upon cGAS–STING axis independent detection. Sensing requires synthesis viral prevented reverse transcriptase inhibitors or active‐site mutation. These demonstrate required can successfully transit avoid sensing.

Язык: Английский

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Spatial resolution of HIV-1 post-entry steps in resting CD4 T cells

Cell Reports, Год журнала: 2024, Номер 43(3), С. 113941 - 113941

Опубликована: Март 1, 2024

Resting CD4 T cells resist productive HIV-1 infection. The HIV-2/simian immunodeficiency virus protein viral accessory X (Vpx) renders these permissive to infection, presumably by alleviating blocks at cytoplasmic reverse transcription and subsequent nuclear import of reverse-transcription/pre-integration complexes (RTC/PICs). Here, spatial analyses using quantitative imaging techniques reveal that capsids containing RTC/PICs are readily imported into the nucleus, recruit host dependency factor CPSF6, translocate speckles in resting cells. Reverse transcription, however, remains incomplete, impeding proviral integration gene expression. Vpx or pharmacological inhibition deoxynucleotide triphosphohydrolase (dNTPase) activity restriction SAM domain HD domain-containing 1 (SAMHD1) increases levels reverse-transcribed cDNA facilitates integration. Nuclear intranuclear transport therefore do not pose important cells, limitation SAMHD1's dNTPase constitutes main pre-integration block

Язык: Английский

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HIV-1 capsid stability and reverse transcription are finely balanced to minimize sensing of reverse transcription products via the cGAS-STING pathway

mBio, Год журнала: 2024, Номер 15(5)

Опубликована: Март 26, 2024

ABSTRACT A critical determinant for early post-entry events, the HIV-1 capsid (CA) protein forms conical core when it rearranges around dimeric RNA genome and associated viral proteins. Although mutations in CA have been reported to alter innate immune sensing of HIV-1, a direct link between stability nucleic acids has not established. Herein, we assessed how manipulating lattice through chemical genetic approaches affects recognition HIV-1. We found that destabilization resulted potent reverse transcription products per se does completely block transcription. Surprisingly, due combined effects enhanced defects nuclear entry, two separate mutants form hyperstable cores induced more potently than destabilizing mutations. At low concentrations allowed accumulation products, CA-targeting compounds GS-CA1 lenacapavir measurably impacted cells modestly HIV. Interestingly, activation observed with viruses containing unstable was abolished by doses lenacapavir. Innate both dependent on cGAS-STING DNA-sensing pathway Overall, our findings demonstrate are finely balanced support minimize cGAS-STING-mediated resulting DNA. IMPORTANCE In particles, proteins enzymes encased proteinaceous composed protein. altering this orthogonal impacts induction responses. Specifically, decreasing results but genomic RNA, cGAS-STING-dependent manner. The recently developed inhibitors Unexpectedly, increased levels cytosolic cDNA, also type I interferon-mediated immunity. Our suggest exposure cytosol host cells.

Язык: Английский

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Lenacapavir disrupts HIV-1 core integrity while stabilizing the capsid lattice

Proceedings of the National Academy of Sciences, Год журнала: 2025, Номер 122(14)

Опубликована: Апрель 1, 2025

Lenacapavir (GS-6207; LEN) is a potent HIV-1 capsid inhibitor approved for treating multidrug-resistant infection. LEN binds to hydrophobic pocket between neighboring (CA) proteins in hexamers and stabilizes the lattice, but its effect on capsids not fully understood. Here, we labeled with green fluorescent protein fused CA (GFP-CA) or fluid-phase GFP content marker (cmGFP) assess LEN’s impact capsids. cores GFP-CA, cmGFP, could be immunostained an anti-GFP antibody were less sensitive capsid-binding host restriction factor MX2, demonstrating that GFP-CA incorporated into lattice stability, whereas cmGFP indicator of core integrity. treatment isolated resulted dose-dependent loss signal while preserving signal, indicating disrupts integrity lattice. In contrast, PF-3450074 (PF74) induced Electron microscopy LEN- PF74-treated viral revealed frequent breakage at narrow end other morphological changes. Our results suggest does prevent nuclear envelope docking inhibits import without PF74 blocks by inhibiting cores, highlighting their different mechanisms inhibition.

Язык: Английский

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