The primary mechanism for highly potent inhibition of HIV-1 maturation by lenacapavir DOI Open Access
Szu‐Wei Huang, Lorenzo Briganti, Arun S. Annamalai

et al.

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

Published: Dec. 7, 2024

Lenacapavir (LEN) is a highly potent, long-acting antiretroviral medication for treating people infected with muti-drug-resistant HIV-1 phenotypes. The inhibitor targets multifaceted functions of the viral capsid protein (CA) during replication. Previous studies have mainly focused on elucidating LEN's mode action ingress. Additionally, has been shown to interfere mature assembly egress. However, mechanism how LEN affects maturation unknown. Here, we show that pharmacologically relevant concentrations do not impair proteolytic processing Gag in virions. Instead, elucidated primary potent inhibition by sub-stoichiometric LEN:CA ratios. exerts opposing effects formation CA pentamers versus hexamers, key capsomere intermediates assembly. impairs pentamers, whereas it induces hexameric lattices imposing an opened conformation and stabilizing dimeric form CA. Consequently, treatment results morphologically atypical virus particles containing malformed, hyper-stable assemblies, which fail infect target cells. Moreover, uncovered inverse correlation between potency levels cell culture assays, accounts ability potently (with pM EC

Language: Английский

HIV-1 capsid shape, orientation, and entropic elasticity regulate translocation into the nuclear pore complex DOI Creative Commons
Arpa Hudait, Gregory A. Voth

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

Citations

22
Pharmacologic hyperstabilisation of the HIV-1 capsid lattice induces capsid failure DOI Creative Commons

K. Faysal,

James Walsh,

Nadine Renner

et al.

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

Citations

19
The capsid lattice engages a bipartite NUP153 motif to mediate nuclear entry of HIV-1 cores DOI Creative Commons
Qi Shen, Sushila Kumari, Chaoyi Xu

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2023, Volume and Issue: 120(13)

Published: March 21, 2023

Increasing evidence has suggested that the HIV-1 capsid enters nucleus in a largely assembled, intact form. However, not much is known about how cone-shaped interacts with nucleoporins (NUPs) nuclear pore for crossing complex. Here, we elucidate NUP153 binds by engaging assembled protein (CA) lattice. A bipartite motif containing both canonical and noncanonical interaction modules was identified at C-terminal tail region of NUP153. The cargo-targeting phenylalanine-glycine (FG) engaged CA hexamer. By contrast, previously unidentified triple-arginine (RRR) targeted tri-hexamer interface capsid. infection studies indicated FG- RRR-motifs were important import cores. Moreover, presence stabilized tubular assemblies vitro. Our results provide molecular-level mechanistic contributes to entry into nucleus.

Language: Английский

Citations

34
HIV-1 Capsid Uncoating Is a Multistep Process That Proceeds through Defect Formation Followed by Disassembly of the Capsid Lattice DOI Creative Commons

Levi B. Gifford,

Gregory B. Melikyan

ACS Nano, Journal Year: 2024, Volume and Issue: 18(4), P. 2928 - 2947

Published: Jan. 19, 2024

The HIV-1 core consists of a cone-shaped capsid shell made protein (CA) hexamers and pentamers encapsulating the viral genome. disassembly, referred to as uncoating, is important for productive infection; however, location, timing, regulation uncoating remain controversial. Here, we employ amber codon suppression directly label CA. In addition, fluid phase fluorescent probe incorporated into detect small defects in lattice. This double-labeling strategy enables visualization single cores vitro living cells, which found always proceed through at least two distinct steps─the formation defect lattice that initiates gradual loss CA below detectable level. Importantly, intact containing markers enter uncoat nucleus, evidenced by sequential both markers, prior establishing infection. two-step process observed different including macrophage line. Notably, lag between release marker terminal appears be independent cell type or reverse transcription much longer (>5-fold) nuclear capsids compared cell-free cytosol, suggesting stabilized capsid-binding factors. Our results imply nucleus initiated localized global

Language: Английский

Citations

11
The primary mechanism for highly potent inhibition of HIV-1 maturation by lenacapavir DOI Creative Commons
Szu‐Wei Huang, Lorenzo Briganti, Arun S. Annamalai

et al.

PLoS Pathogens, Journal Year: 2025, Volume and Issue: 21(1), P. e1012862 - e1012862

Published: Jan. 27, 2025

Lenacapavir (LEN) is a highly potent, long-acting antiretroviral medication for treating people infected with muti-drug-resistant HIV-1 phenotypes. The inhibitor targets multifaceted functions of the viral capsid protein (CA) during replication. Previous studies have mainly focused on elucidating LEN’s mode action ingress. Additionally, has been shown to interfere mature assembly egress. However, mechanism how LEN affects maturation unknown. Here, we show that pharmacologically relevant concentrations do not impair proteolytic processing Gag in virions. Instead, elucidated primary potent inhibition by sub-stoichiometric LEN:CA ratios. exerts opposing effects formation CA pentamers versus hexamers, key capsomere intermediates assembly. impairs pentamers, whereas it induces hexameric lattices imposing an opened conformation and stabilizing dimeric form CA. Consequently, treatment results morphologically atypical virus particles containing malformed, hyper-stable assemblies, which fail infect target cells. Moreover, uncovered inverse correlation between potency levels cell culture assays, accounts ability potently (with picomolar EC 50 values) inhibit at clinically drug concentrations.

Language: Английский

Citations

1
Lenacapavir disrupts HIV-1 core integrity while stabilizing the capsid lattice DOI Creative Commons
Chenglei Li, Ryan C. Burdick,

Rokeya Siddiqui

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2025, Volume and Issue: 122(14)

Published: April 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.

Language: Английский

Citations

1
HIV capsids: orchestrators of innate immune evasion, pathogenesis and pandemicity DOI
Kate L. Morling, Mohamed ElGhazaly,

R Milne

et al.

Journal of General Virology, Journal Year: 2025, Volume and Issue: 106(1)

Published: Jan. 13, 2025

Human immunodeficiency virus (HIV) is an exemplar virus, still the most studied and best understood a model for mechanisms of viral replication, immune evasion pathogenesis. In this review, we consider earliest stages HIV infection from transport virion contents through cytoplasm to integration genome into host chromatin. We present holistic virus-host interaction during pivotal stage infection. Central process capsid. The last 10 years have seen transformation in way understand capsid structure function. review key discoveries our latest thoughts on as dynamic regulator innate chromatin targeting. also accessory proteins Vpr Vpx because they are incorporated particles where collaborate with capsids manipulate defensive cellular responses argue that effective regulation uncoating immunity define pandemic potential pathogenesis, how comparison different lineages can reveal what makes lentiviruses special.

Language: Английский

Citations

0
Cell-free assays reveal that the HIV-1 capsid protects reverse transcripts from cGAS immune sensing DOI Creative Commons
Tiana M. Scott,

Lydia M. Arnold,

Jordan A. Powers

et al.

PLoS Pathogens, Journal Year: 2025, Volume and Issue: 21(1), P. e1012206 - e1012206

Published: Jan. 28, 2025

Retroviruses can be detected by the innate immune sensor cyclic GMP-AMP synthase (cGAS), which recognizes reverse-transcribed DNA and activates an antiviral response. However, extent to HIV-1 shields its genome from cGAS recognition remains unclear. To study this process in mechanistic detail, we reconstituted reverse transcription, release, sensing of a cell-free system. We found that wild-type capsids protect viral genomes even after completing transcription. Viral could “deprotected” thermal stress, capsid mutations, or reduced concentrations inositol hexakisphosphate (IP6) destabilize capsid. Strikingly, inhibitor lenacapavir also disrupted cores dramatically potentiated activity, both vitro cellular infections. Our results provide biochemical evidence lattice conceals chemical physical disruption core expose activate signaling.

Language: Английский

Citations

0
Considerations for capsid-targeting antiretrovirals in pre-exposure prophylaxis DOI
William M. McFadden,

Mia Færch,

Karen A. Kirby

et al.

Trends in Molecular Medicine, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

Citations

0
Structural and mechanistic bases for resistance of the M66I capsid variant to lenacapavir DOI Creative Commons
Lorenzo Briganti, Arun S. Annamalai, S.M. Bester

et al.

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

Published: April 15, 2025

ABSTRACT Lenacapavir (LEN) is the first-in-class viral capsid protein (CA) targeting antiretroviral for treating multi-drug-resistant HIV-1 infection. Clinical trials and cell culture experiments have identified resistance-associated mutations (RAMs) in vicinity of hydrophobic CA pocket targeted by LEN. The M66I substitution conferred far highest level resistance to inhibitor compared other RAMs. Here we investigated structural mechanistic bases how change affects LEN binding replication. high-resolution X-ray structure CA(M66I) hexamer revealed that β-branched side chain Ile66 induces steric hindrance specifically LEN, thereby markedly reducing affinity. By contrast, did not affect Phe-Gly (FG)-motif-containing cellular cofactors CPSF6, NUP153, or SEC24C, which engage same CA. In culture, variant acquire compensatory mutations. Analysis replication intermediates (M66I CA) predominantly formed correctly matured cores, were more stable than their wild-type counterparts. mutant cores stably bound nuclear envelope but failed penetrate inside nucleus. Furthermore, altered integration targeting. Taken together, our findings elucidate insights into confers remarkable Moreover, provide a powerful means future medicinal chemistry efforts rationally develop second-generation inhibitors with higher barrier resistance. IMPORTANCE highly potent long-acting works unique mechanism protein. used combination antiretrovirals treat infection heavily treatment-experienced adults. clinical preexposure prophylaxis (PrEP) interim results indicating 100% efficacy prevent infections. However, one notable shortcoming relatively low emergent near site on capsid. was most prevalent patients receiving studies described here underlying marked inhibitor. will aid next generation enhanced barriers

Language: Английский

Citations

0