Cone-shaped HIV-1 capsids are transported through intact nuclear pores

Cell, Journal Year: 2021, Volume and Issue: 184(4), P. 1032 - 1046.e18

Published: Feb. 1, 2021

Human immunodeficiency virus (HIV-1) remains a major health threat. Viral capsid uncoating and nuclear import of the viral genome are critical for productive infection. The size HIV-1 is generally believed to exceed diameter pore complex (NPC), indicating that has occur prior import. Here, we combined correlative light electron microscopy with subtomogram averaging capture structural status reverse transcription-competent complexes in infected T cells. We demonstrated NPC cellulo sufficient apparently intact, cone-shaped capsids. Subsequent import, detected disrupted empty fragments, replication occurs by breaking open, not disassembly into individual subunits. Our data directly visualize key step enhance our mechanistic understanding life cycle.

Language: Английский

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HIV-1 uncoats in the nucleus near sites of integration

Proceedings of the National Academy of Sciences, Journal Year: 2020, Volume and Issue: 117(10), P. 5486 - 5493

Published: Feb. 24, 2020

Significance For several decades, retroviral core uncoating has been thought to occur in the cytoplasm coordination with reverse transcription, and while some recent studies have concluded that HIV-1 occurs at nuclear envelope during import, none nucleus. Here, we developed methods study by direct labeling quantification of viral capsid protein associated infectious cores produced transcriptionally active proviruses. We find nuclei infected cells are largely intact uncoat near their integration sites just before integration. These unexpected findings fundamentally change our understanding postentry replication events.

Language: Английский

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Nuclear pore blockade reveals that HIV-1 completes reverse transcription and uncoating in the nucleus

Nature Microbiology, Journal Year: 2020, Volume and Issue: 5(9), P. 1088 - 1095

Published: June 1, 2020

Language: Английский

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Structural and mechanistic bases for a potent HIV-1 capsid inhibitor

Science, Journal Year: 2020, Volume and Issue: 370(6514), P. 360 - 364

Published: Oct. 16, 2020

The potent HIV-1 capsid inhibitor GS-6207 is an investigational principal component of long-acting antiretroviral therapy. We found that inhibits by stabilizing and thereby preventing functional disassembly the shell in infected cells. X-ray crystallography, cryo-electron microscopy, hydrogen-deuterium exchange experiments revealed tightly binds two adjoining subunits promotes distal intra- inter-hexamer interactions stabilize curved lattice. In addition, interferes with binding to cellular cofactors Nup153 CPSF6 mediate viral nuclear import direct integration into gene-rich regions chromatin. These findings elucidate structural insights multimodal, antiviral activity provide a means for rationally developing second-generation therapies.

Language: Английский

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HIV-1 cores retain their integrity until minutes before uncoating in the nucleus

Proceedings of the National Academy of Sciences, Journal Year: 2021, Volume and Issue: 118(10)

Published: March 1, 2021

Significance Here, we used a fluorescent protein that is free in solution and trapped nuclear HIV-1 capsids to demonstrate the retain integrity prevent mixing of macromolecules within viral core cellular environment until just before integration. We also found capsid maintained minutes disassembly nucleus, revealing uncoating proceeds rapidly after loss. These valuable insights into early stage replication indicate intact are imported through pores, reverse transcription mostly completed capsids, preintegration complex-host interactions facilitating integration target site selection must occur short time frame between

Language: Английский

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HIV-1 uncoating by release of viral cDNA from capsid-like structures in the nucleus of infected cells

eLife, Journal Year: 2021, Volume and Issue: 10

Published: April 27, 2021

HIV-1 replication commences inside the cone-shaped viral capsid, but timing, localization, and mechanism of uncoating are under debate. We adapted a strategy to visualize individual reverse-transcribed cDNA molecules their association with cellular proteins using fluorescence correlative-light-and-electron-microscopy (CLEM). specifically detected nuclei, not in cytoplasm. Nuclear initially co-localized fluorescent integrase fusion (IN-FP) CA (capsid) protein, cDNA-punctae separated from IN-FP/CA over time. This phenotype was conserved primary target cells, nuclear complexes exhibiting strong CA-signals all cell types. CLEM revealed capsid-like structures apparently broken capsid-remnants at position IN-FP signals elongated chromatin-like punctae lacking IN-FP. Our data argue for by physical disruption rather than cooperative disassembly CA-lattice, followed separation pre-integration complex.

Language: Английский

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The HIV capsid mimics karyopherin engagement of FG-nucleoporins

Nature, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 24, 2024

Abstract HIV can infect non-dividing cells because the viral capsid overcome selective barrier of nuclear pore complex and deliver genome directly into nucleus 1,2 . Remarkably, intact is more than 1,000 times larger size limit prescribed by diffusion 3 This in central channel composed intrinsically disordered nucleoporin domains enriched phenylalanine–glycine (FG) dipeptides. Through multivalent FG interactions, cellular karyopherins their bound cargoes solubilize this phase to drive nucleocytoplasmic transport 4 By performing an vitro dissection complex, we show that a pocket on surface similarly interacts with motifs from multiple nucleoporins interaction licences capsids penetrate FG-nucleoporin condensates. karyopherin mimicry model addresses key conceptual challenge for role entry offers explanation as how exogenous entity much any known cargo may be able non-destructively breach envelope.

Language: Английский

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HIV-1 capsids enter the FG phase of nuclear pores like a transport receptor

Nature, Journal Year: 2024, Volume and Issue: 626(8000), P. 843 - 851

Published: Jan. 24, 2024

Abstract HIV-1 infection requires nuclear entry of the viral genome. Previous evidence suggests that this proceeds through pore complexes (NPCs), with 120 × 60 nm capsid squeezing an approximately 60-nm-wide central channel 1 and crossing permeability barrier NPC. This can be described as FG phase 2 is assembled from cohesively interacting phenylalanine–glycine (FG) repeats 3 selectively permeable to cargo captured by transport receptors (NTRs). Here we show assemblies target NPCs efficiently in NTR-independent manner bind directly several types repeats, including barrier-forming cohesive repeats. Like NTRs, readily partitions into vitro serve NPC mimic excludes much smaller inert probes such mCherry. Indeed, protein greatly enhanced assembly, which also allows encapsulated clients enter. Thus, our data indicate behaves like NTR, its interior serving a container. Because capsid-coating trans -acting NTRs would increase diameter 10 or more, suggest ‘self-translocating’ undermines size restrictions imposed scaffold, thereby bypassing otherwise effective infection.

Language: Английский

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HIV-1 capsid shape, orientation, and entropic elasticity regulate translocation into the nuclear pore complex

Proceedings of the National Academy of Sciences, Journal Year: 2024, Volume and Issue: 121(4)

Published: Jan. 19, 2024

Nuclear import and uncoating of the viral capsid are critical steps in HIV-1 life cycle that serve to transport release genomic material into nucleus. Viral core involves translocating at nuclear pore complex (NPC). Notably, central channel NPC appears often accommodate allow passage intact capsid, though mechanistic details process remain be fully understood. Here, we investigate molecular interactions operate concert between regulate translocation through channel. To this end, develop a “bottom-up” coarse-grained (CG) model human from recently released cryo-electron tomography structure then construct composite membrane-embedded CG models. We find successful cytoplasmic side is contingent on compatibility morphology dimension proper orientation approach side. The dynamics driven by maximizing contacts phenylalanine-glycine nucleoporins capsid. For docked capsids, structural analysis reveals correlated striated patterns lattice disorder likely related intrinsic elasticity. Uncondensed inside augments overall Our results suggest “elasticity” can also aid adapt stress structurally during translocation.

Language: Английский

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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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