Structural determinants of spike infectivity in bat SARS-like coronaviruses RsSHC014 and WIV1

Journal of Virology, Год журнала: 2024, Номер 98(8)

Опубликована: Июль 19, 2024

The recurrent spillovers of coronaviruses (CoVs) have posed severe threats to public health and the global economy. Bat acute respiratory syndrome (SARS)-like CoVs RsSHC014 WIV1, currently circulating in bat populations, are poised for human emergence. trimeric spike (S) glycoprotein, responsible receptor recognition membrane fusion, plays a critical role cross-species transmission infection. Here, we determined cryo-electron microscopy (EM) structures S protein closed state at 2.9 Å, WIV1 2.8 intermediate 4.0 Å. In state, one receptor-binding domain (RBD) is "down" position, while other two RBDs exhibit poor density. We also resolved complex structure bound ACE2 (hACE2) 4.5 which provides structural basis future emergence humans. Through biochemical experiments, found that despite strong binding affinities between both civet ACE2, pseudoviruses RsSHC014, but not failed infect 293T cells overexpressing either or ACE2. Mutagenesis analysis revealed Y623H substitution, located SD2 region, significantly improved cell entry efficiency pseudoviruses, likely accomplished by promoting open conformation glycoproteins. Our findings emphasize necessity efficient RBD lifting tight RBD-hACE2 viral infection underscore significance 623 site glycoprotein infectivity SARS-like CoVs.

Язык: Английский

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Molecular Insights into Cross-Species Spillover of Coronavirus HKU5 via ACE2 Receptor Recognition

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Янв. 10, 2025

Abstract Coronaviruses represent a significant zoonotic threat, with host adaptation serving as pivotal determinant of cross-species transmission. The bat-derived β-coronavirus HKU5 utilizes its spike (S) protein for receptor recognition and viral entry. Here, we report the cryo-electron microscopy (cryo-EM) structure S in closed conformation. Two fatty acids are bound to protein, stabilizing Furthermore, solve receptor-binding domain (RBD) complex peptidase (PD) Pipistrellus abramus angiotensin-converting enzyme 2 (ACE2), uncovering unique binding mode distinct from other coronaviruses that use ACE2 their receptor. Evolutionary functional analyses indicate mutations on RBD can modulate binding, while conservation structural modeling suggest has potential cross species barrier. Notably, identify orthologs avian species, such Pitta sordida , support stable interaction. These findings provide molecular insights into underscore importance surveillance this virus risk.

Язык: Английский

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Diketopiperazine/piperidine alkaloid as a potential broad-spectrum coronaviral entry inhibitor identified by supercomputer-based virtual screening from a large natural product-based library

Biomedicine & Pharmacotherapy, Год журнала: 2025, Номер 183, С. 117841 - 117841

Опубликована: Янв. 13, 2025

Язык: Английский

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Human HKU1-reactive CD4 T cells are enriched for cytolytic potential that persists in older adults

The Journal of Infectious Diseases, Год журнала: 2025, Номер unknown

Опубликована: Янв. 15, 2025

The emergence of SARS-CoV-2 increased interest in cellular immunity established by infections with human coronaviruses (HCoVs). Using PBMC from a cohort subjects collected prior to 2019, we assessed the abundance and phenotype these CD4 T cells using cytokine Elispot assays. Unexpectedly, cytotoxic potential was uniquely enriched amongst HKU1-reactive cells, as measured quantification granzyme producing cells. Also, although dramatic losses HCoV-specific for OC43, NL63 229E-specific were observed older relative younger adults, exhibited minimal age-dependent differences this phenotype.

Язык: Английский

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CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex

Scientific Reports, Год журнала: 2025, Номер 15(1)

Опубликована: Апрель 7, 2025

Abstract The TMPRSS2 cell surface protease is used by a broad range of respiratory viruses to facilitate entry into target cells. Together with ACE2, represents key factor for SARS-CoV-2 infection, as mediates cleavage viral spike protein, enabling direct fusion the envelope host membrane. Since start COVID-19 pandemic, has gained attention therapeutic inhibitors which would inhibit but little known about regulation, particularly in types physiologically relevant infection. Here, we performed an unbiased genome-wide CRISPR-Cas9 library screen, together targeted at epigenetic modifiers and transcriptional regulators, identify cellular factors that modulate expression human colon epithelial We find endogenous regulated Elongin BC-VHL complex HIF transcription factors. Depletion B or treatment cells PHD resulted downregulation inhibition show still utilised Omicron variants colonic Our study enhances our understanding regulation

Язык: Английский

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Structural basis for the interaction between human coronavirus HKU1 spike receptor binding domain and its receptor TMPRSS2

Cell Discovery, Год журнала: 2024, Номер 10(1)

Опубликована: Авг. 8, 2024

Язык: Английский

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Identification of the critical residues of TMPRSS2 for entry and host range of human coronavirus HKU1

Journal of Virology, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 11, 2024

Human coronavirus (CoV) HKU1 infection typically causes common cold but can lead to pneumonia in children, older people, and immunosuppressed individuals. Recently, human transmembrane serine protease 2 (hTMPRSS2) was identified as the functional receptor for HKU1, its region residues critical S binding remain elusive. In this study, we find that could utilize hamster, not rat, mouse, or bat TMPRSS2 virus entry, displaying a narrow host range. Using human-bat chimeras, show peptidase (SP) domain of is essential entry HKU1. Further extensive mutagenesis analyses C-terminal regions SP domains TMPRSS2s identify 417 469 Replacement either D417 Y469 with asparagine hTMPRSS2 abolishes abilities mediate pseudovirions cell-cell fusion, whereas substitution N417 D N469 Y (bTMPRSS2) renders it supporting entry. Our findings contribute deeper understanding coronavirus-receptor interactions cross-species transmission.IMPORTANCEThe proteins their cognate receptors determine range transmission potential. found be Here, bat, serve Moreover, swapping at positions bTMPRSS2 corresponding confers pseudovirions, indicating role these study responsible interaction

Язык: Английский

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TMPRSS2 in microbial interactions: Insights from HKU1 and TcsH

PLoS Pathogens, Год журнала: 2024, Номер 20(11), С. e1012677 - e1012677

Опубликована: Ноя. 20, 2024

Transmembrane Serine Protease 2 (TMPRSS2), known primarily for its role as a protease, has emerged critical receptor microbial agents such human coronavirus HKU1 and exotoxin TcsH. utilizes both sialoglycan TMPRSS2 cellular entry, where primes the spike protein binding. undergoes autocleavage to enhance affinity spike, facilitating viral membrane fusion postcleavage. Interestingly, TMPRSS2’s catalytic function is dispensable TcsH interactions, suggesting alternative roles in pathogenesis. Structural insights highlight potential therapeutic targets against infections cancers, leveraging interactions drug development. Understanding interplay between microbes opens new avenues targeting developing treatments infections.

Язык: Английский

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High-resolution crystal structure of human coronavirus HKU1 receptor binding domain bound to TMPRSS2 receptor

hLife, Год журнала: 2024, Номер unknown

Опубликована: Окт. 1, 2024

Язык: Английский

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Identification of nitrile-containing isoquinoline-related natural product derivatives as coronavirus entry inhibitors in silico and in vitro

Biomedicine & Pharmacotherapy, Год журнала: 2024, Номер 180, С. 117517 - 117517

Опубликована: Окт. 1, 2024

Язык: Английский

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