Pharmacologic hyperstabilisation of the HIV-1 capsid lattice induces capsid failure

eLife, Journal Year: 2024, Volume and Issue: 13

Published: Feb. 13, 2024

The HIV-1 capsid has emerged as a tractable target for antiretroviral therapy. Lenacapavir, developed by Gilead Sciences, is the first capsid-targeting drug approved medical use. Here, we investigate effect of lenacapavir on HIV stability and uncoating. We employ single particle approach that simultaneously measures content release lattice persistence. demonstrate lenacapavir's potent antiviral activity predominantly due to lethal hyperstabilisation resultant loss compartmentalisation. This study highlights disrupting metastability powerful strategy development novel antivirals.

Language: Английский

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Passage of the HIV capsid cracks the nuclear pore

Cell, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Language: Английский

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Elasticity of the HIV-1 core facilitates nuclear entry and infection

PLoS Pathogens, Journal Year: 2024, Volume and Issue: 20(9), P. e1012537 - e1012537

Published: Sept. 11, 2024

HIV-1 infection requires passage of the viral core through nuclear pore cell, a process that depends on functions capsid. Recent studies have shown cores enter nucleus prior to capsid disassembly. Interactions with complex are necessary but not sufficient for entry, and mechanism by which traverses comparably sized is unknown. Here we show highly elastic this property linked entry infectivity. Using atomic force microscopy-based approaches, found purified wild type rapidly returned their normal conical morphology following severe compression. Results from independently performed molecular dynamic simulations mature also revealed its property. Analysis four mutants exhibit impaired mutant brittle. Adaptation two viruses in cell culture resulted additional substitutions restored elasticity rescued infectivity entry. We capsid-targeting compound PF74 antiviral drug Lenacepavir reduce block at concentrations preserve interactions between envelope. Our results indicate fundamental enables thereby facilitating infection. These provide new insights into role mechanisms inhibitors.

Language: Английский

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Passage of the HIV capsid cracks the nuclear pore

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 23, 2024

Summary Upon infection, human immunodeficiency virus (HIV-1) releases its cone-shaped capsid into the cytoplasm of infected T-cells and macrophages. As largest known cargo, enters nuclear pore complex (NPC), driven by interactions with numerous FG-repeat nucleoporins (FG-Nups). Whether NPCs structurally adapt to passage whether capsids are modified during remains unknown, however. Here, we combined super-resolution correlative microscopy cryo electron tomography molecular simulations study entry HIV-1 in primary We found that cytosolically bound cyclophilin A is stripped off entering NPC, hexagonal lattice largely intact inside beyond central channel. Strikingly, NPC scaffold rings frequently crack passage, consistent computer indicating need for widening. The unique cone shape facilitates helps their rings.

Language: Английский

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HIV-1 usurps mixed-charge domain-dependent CPSF6 phase separation for higher-order capsid binding, nuclear entry and viral DNA integration

Nucleic Acids Research, Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 25, 2024

Abstract HIV-1 integration favors nuclear speckle (NS)-proximal chromatin and viral infection induces the formation of capsid-dependent CPSF6 condensates that colocalize with speckles (NSs). Although displays liquid-liquid phase separation (LLPS) activity in vitro, contributions its different intrinsically disordered regions, which includes a central prion-like domain (PrLD) capsid binding FG motif C-terminal mixed-charge (MCD), to LLPS remain unclear. Herein, we determined PrLD MCD both contribute vitro. Akin mutant CPSF6, cells expressing MCD-deleted uncharacteristically arrested at rim. While heterologous MCDs effectively substituted for function during infection, Arg-Ser domains from related SR proteins were largely ineffective. wildtype displayed similar affinities, imparted LLPS-dependent higher-order co-aggregation capsids vitro cellulo. NS depletion reduced puncta without significantly affecting into NS-proximal chromatin, appending onto protein partially restored virus penetration targeting knockout cells. We conclude MCD-dependent condensation underlies post-nuclear incursion DNA pathogenesis.

Language: Английский

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Interplay between the cyclophilin homology domain of RANBP2 and MX2 regulates HIV-1 capsid dependencies on nucleoporins

mBio, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 24, 2025

ABSTRACT Interlinked interactions between the viral capsid (CA), nucleoporins (Nups), and antiviral protein myxovirus resistance 2 (MX2/MXB) influence human immunodeficiency virus 1 (HIV-1) nuclear entry outcome of infection. Although RANBP2/NUP358 has been repeatedly identified as a critical player in HIV-1 import MX2 activity, mechanism by which RANBP2 facilitates infection is not well understood. To explore MX2, CA, RANBP2, we utilized CRISPR-Cas9 to generate cell lines expressing from its endogenous locus but lacking C-terminal cyclophilin (Cyp) homology domain found that both HIV-2 infections were reduced significantly ΔCyp cells. Importantly, although still localized pore complex cells, activity against was decreased. By generating cells specific point mutations RANBP2-Cyp domain, determined effect on anti-HIV-1 due direct CA. We further CypA have similar effects integration targeting. Finally, Nup requirements for HIV altered domain. These findings demonstrate affects sensitivity altering CA-specific with cellular factors affect IMPORTANCE Human into nucleus an essential step replication involves (CA) multiple proteins, including (Nups) such RANBP2. Nups also mediate function (MX2); however, determining precise role proved challenging nature (NPC) significant pleiotropic elicited depletion. used gene editing assess activity. find this infection, nucleoporin requirements, sensitivity, targeting manner, providing detailed insights how contributes

Language: Английский

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Conformational dynamics of the nuclear pore complex central channel

Biochemical Society Transactions, Journal Year: 2025, Volume and Issue: 53(01)

Published: Feb. 7, 2025

The nuclear pore complex (NPC) is a vital regulator of molecular transport between the nucleus and cytoplasm in eukaryotic cells. At heart NPC’s function are intrinsically disordered phenylalanineglycine-rich nucleoporins (FG-Nups), which form dynamic permeability barrier within central channel. This nature facilitates efficient nucleocytoplasmic but also poses significant challenges to its characterization, especially nano-confined environment NPC. Recent advances experimental techniques, such as cryo-electron microscopy, atomic force fluorescence magnetic resonance, along with computational modeling, have illuminated conformational flexibility FG-Nups, underpins their functional versatility. review synthesizes these advancements, emphasizing how disruptions FG-Nup behavior—caused by mutations or pathological interactions—contribute diseases neurodegenerative disorders, aging-related decline, viral infections. Despite progress, persist deciphering dynamics crowded cellular environment, under conditions. Addressing gaps critical for advancing therapeutic strategies targeting NPC dysfunction disease progression.

Language: Английский

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In-cell structure and snapshots of copia retrotransposons in intact tissue by cryoelectron tomography

Cell, Journal Year: 2025, Volume and Issue: unknown

Published: March 1, 2025

Long terminal repeat (LTR) retrotransposons belong to the transposable elements (TEs), autonomously replicating genetic that integrate into host's genome. Among animals, Drosophila melanogaster serves as an important model organism for TE research and contains several LTR retrotransposons, including Ty1-copia family, which is evolutionarily related retroviruses forms virus-like particles (VLPs). In this study, we use cryo-focused ion beam (FIB) milling lift-out approaches visualize copia VLPs in ovarian cells intact egg chambers, resolving situ capsid structure 7.7 Å resolution by cryoelectron tomography (cryo-ET). Although cytoplasmic vary size, nuclear are homogeneous form densely packed clusters, supporting a import acts size selector. Analyzing flies deficient TE-suppressing PIWI-interacting RNA (piRNA) pathway, observe copia's translocation nucleus during spermatogenesis. Our findings provide insights replication cycle cellular structural biology of active retrotransposon.

Language: 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, Journal Year: 2024, Volume and Issue: 15(5)

Published: March 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.

Language: Английский

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Spatiotemporal binding of cyclophilin A and CPSF6 to capsid regulates HIV-1 nuclear entry and integration

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 9, 2024

Abstract Human immunodeficiency virus type 1 (HIV-1) capsid, which is the target of antiviral lenacapavir, protects viral genome and binds multiple host proteins to influence intracellular trafficking, nuclear import, integration. Previously, we showed that capsid binding cleavage polyadenylation specificity factor 6 (CPSF6) in cytoplasm competitively inhibited by cyclophilin A (CypA) regulates infection. Here determined a mutant with increased CypA affinity had significantly reduced entry mislocalized However, disruption restored entry, integration, infection CPSF6-dependent manner. Furthermore, relocalization expression from cell nucleus failed restore HIV-1 Our results clarify sequential CPSF6 required for optimal integration targeting, informing antiretroviral therapies contain lenacapavir.

Language: Английский

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