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

Passage of the HIV capsid cracks the nuclear pore DOI Creative Commons
Jan Philipp Kreysing, Maziar Heidari, Vojtěch Žíla

et al.

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

Published: Jan. 1, 2025

Language: Английский

Citations

8
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
HIV-1 capsid stability and reverse transcription are finely balanced to minimize sensing of reverse transcription products via the cGAS-STING pathway DOI Creative Commons
Jenna E. Eschbach, Maritza Puray‐Chavez, Shawn Mohammed

et al.

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

Citations

8
HIV-1 usurps mixed-charge domain-dependent CPSF6 phase separation for higher-order capsid binding, nuclear entry and viral DNA integration DOI Creative Commons
Sooin Jang, Gregory J. Bedwell, Satya P. Singh

et al.

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

Citations

8
Strategies for the Viral Exploitation of Nuclear Pore Transport Pathways DOI Creative Commons
Xin Zhang, Keesiang Lim,

Yujia Qiu

et al.

Viruses, Journal Year: 2025, Volume and Issue: 17(2), P. 151 - 151

Published: Jan. 23, 2025

Viruses frequently exploit the host’s nucleocytoplasmic trafficking machinery to facilitate their replication and evade immune defenses. By encoding specialized proteins other components, they strategically target host nuclear transport receptors (NTRs) nucleoporins within spiderweb-like inner channel of pore complex (NPC), enabling efficient access nucleus. This review explores intricate mechanisms governing import export viral with a focus on interplay between factors determinants that are essential for these processes. Given pivotal role shuttling in life cycle, we also examine therapeutic strategies aimed at disrupting pathways. includes evaluating efficacy pharmacological inhibitors impairing assessing potential as antiviral treatments. Furthermore, emphasize need continued research develop targeted therapies leverage vulnerabilities trafficking. Emerging high-resolution techniques, such advanced imaging computational modeling, transforming our understanding dynamic interactions viruses NPC. These cutting-edge tools driving progress identifying novel opportunities uncovering deeper insights into pathogenesis. highlights importance advancements paving way innovative strategies.

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
CPSF6 Promotes HIV-1 Preintegration Complex Function DOI Creative Commons
Evan Chaudhuri, Sooin Jang, Rajasree Chakraborty

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 29, 2025

ABSTRACT Cleavage and polyadenylation specificity factor 6 (CPSF6) is part of the cellular cleavage I mammalian (CFIm) complex that regulates mRNA processing polyadenylation. CPSF6 also functions as a HIV-1 capsid (CA) binding host promotes viral DNA integration targeting into gene dense regions genome. However, effects on activity preintegration (PIC) - machinery carries out to establish infection unknown. To study CPSF6’s role in PIC function, we extracted PICs from cells depleted or expressing mutant cannot bind CA. These exhibited significantly lower when compared control PICs. Addition recombinant restored cells, suggesting direct function. solidify effect inoculated CPSF6-depleted CPSF6-mutant with particles measured A significant reduction these was detected this defect not consequence reduced reverse transcription nuclear entry. Additionally, viruses deficient CA-CPSF6 showed no cells. Finally, sequencing analysis revealed redirected Collectively, results suggest CPSF6-CA interaction function both vitro infected IMPORTANCE dependent virus factors. molecular details virus-host interactions are fully understood. For instance, provides interfaces for several one such capsid-binding factor, whose regulate Initial work identified truncated cytosolic form restricted HIV by blocking it now established full-length primarily Here report complexes (PICs). We observed disruption target directed away gene-dense regions. findings demonstrate critical targeting.

Language: Английский

Citations

0
Structural basis for HIV-1 capsid adaption to rescue IP6-packaging deficiency DOI Creative Commons
Yanan Zhu, Alex B. Kleinpeter, Juan S. Rey

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 9, 2025

Abstract Inositol hexakisphosphate (IP6) promotes HIV-1 assembly via its interaction with the immature Gag lattice, effectively enriching IP6 within virions. During particle maturation, protease cleaves polyproteins comprising releasing from original binding site and liberating capsid (CA) domain of Gag. then mature CA protein into shell viral core, which is required for infection new target cells. Recently, we reported mutants that assemble virions independently IP6. However, these are non-infectious unable to stable capsids. Here, identified a mutation in C-terminus – G225R restores formation infectivity IP6-packaging-deficient mutants. Furthermore, show facilitates vitro purified capsid-like particles (CLPs) at concentrations well below those WT CLP assembly. Using single-particle cryoEM, solved structures hexamer hexameric lattice CLPs harbouring assembled low-IP6 conditions. The high-resolution (2.7 Å) cryoEM structure combined molecular dynamics simulations revealed otherwise flexible disordered becomes structured, extending pseudo two-fold hexamer-hexamer interface, thereby stabilizing capsid. This work uncovers structural mechanism by adapts deficiency packaging. ability promote conditions provides valuable tool capsid-related studies may indicate heretofore unknown role unstructured

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
Correlative In Situ Cryo-ET Reveals Cellular and Viral Remodeling Associated with Selective HIV-1 Core Nuclear Import DOI Creative Commons

Zhen Hou,

Y. Shen, Stanley Fronik

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: March 4, 2025

Abstract Lentiviruses like HIV-1 infect non-dividing cells by traversing the nuclear pore, but studying this process has been challenging due to its scarcity and dynamic nature in infected cells. Here, we developed a robust cell-permeabilization system that recapitulates import established an integrated cryo-correlative workflow combining cryo-CLEM, cryo-FIB, cryo-ET for targeted imaging of process. These advancements enabled successful capture 1,899 cores at various stages import. Statistical structural analyses native wild-type mutant revealed depends on both capsid elasticity pore adaptability, as well factors such CPSF6. Brittle fail enter complex (NPC), while CPSF6-binding-deficient stall inside NPC, resulting impaired Intriguingly, pores function selective filters favoring smaller, tube-shaped cores. Our study opens new avenues dissecting biochemistry biology downstream events including core uncoating potentially integration, with unprecedented detail.

Language: Английский

Citations

0