Human coronaviruses: activation and antagonism of innate immune responses DOI
Nikhila S. Tanneti, Helen A. Stillwell, Susan R. Weiss

et al.

Microbiology and Molecular Biology Reviews, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 19, 2024

SUMMARY Human coronaviruses cause a range of respiratory diseases, from the common cold (HCoV-229E, HCoV-NL63, HCoV-OC43, and SARS-CoV-2) to lethal pneumonia (SARS-CoV, SARS-CoV-2, MERS-CoV). Coronavirus interactions with host innate immune antiviral responses are an important determinant disease outcome. This review compares host’s response different human coronaviruses. Host defenses discussed in this include frontline against viruses nasal epithelium, early sensing viral infection by effectors, double-stranded RNA stress-induced pathways, antagonism conferred conserved coronavirus nonstructural proteins genus-specific accessory proteins. The HCoV-229E -NL63 induce robust interferon signaling related SARS-CoV SARS-CoV-2 intermediate levels activation, MERS-CoV shuts down these pathways almost completely.

Language: Английский

A guanidine-based coronavirus replication inhibitor which targets the nsp15 endoribonuclease and selects for interferon-susceptible mutant viruses DOI Creative Commons
Benjamin Van Loy, Eugènia Pujol, Kenichi Kamata

et al.

PLoS Pathogens, Journal Year: 2025, Volume and Issue: 21(2), P. e1012571 - e1012571

Published: Feb. 11, 2025

The approval of COVID-19 vaccines and antiviral drugs has been crucial to end the global health crisis caused by SARS-CoV-2. However, prepare for future outbreaks from drug-resistant variants novel zoonotic coronaviruses (CoVs), additional therapeutics with a distinct mechanism are needed. Here, we report guanidine-substituted diphenylurea compound that suppresses CoV replication interfering uridine-specific endoribonuclease (EndoU) activity viral non-structural protein-15 (nsp15). This compound, designated EPB-113, exhibits strong selective cell culture against human coronavirus 229E (HCoV-229E) also Viruses, selected under EPB-113 pressure, carried resistance sites at or near catalytic His250 residue nsp15-EndoU domain. Although best-known function EndoU is avoid induction type I interferon (IFN-I) lowering levels dsRNA, was found mainly act via an IFN-independent mechanism, situated during RNA synthesis. Using combination biophysical enzymatic assays recombinant nsp15 proteins HCoV-229E SARS-CoV-2, discovered enhances cleavage hexameric nsp15, while reducing its thermal stability. explains why virus escapes acquiring site mutations which impair binding abolish activity. Since EPB-113-resistant mutant viruses induce high IFN-I effectors, they proved unable replicate in macrophages were readily outcompeted wild-type upon co-infection fibroblast cells. Our findings suggest targeting can be achieved molecule induces conformational change this protein, resulting higher impairment Based on appealing profile conclude challenging but highly relevant drug target.

Language: Английский

Citations

0
Natural Target‐Mimicking Translocation‐Based Fluorescent Sensor for Detection of SARS‐CoV‐2 PLpro Protease Activity and Virus Infection in Living Cells DOI Open Access
Elena L. Sokolinskaya, Olga N. Ivanova,

Irina T. Fedyakina

et al.

Published: May 14, 2024

Papain-like protease PLpro, a domain within large polyfunctional protein nsp3, plays key roles in the life cycle of SARS-CoV-2, being responsible for first events cleavage polyprotein into individual proteins (nsp1-4) as well suppression cellular immunity. Here, we developed new genetically encoded fluorescent sensor, named PLpro-ERNuc, detection PLpro activity living cells using translocation-based readout. The sensor was designed follows. A fragment nsp3 used to direct on cytoplasmic surface endoplasmic reticulum (ER) membrane, thus closely mimicking natural target PLpro. part included two bright - red mScarlet I and green mNeonGreen, separated by linker with site. Nuclear localization signal (NLS) attached ensure accumulation mNeonGreen nucleus upon cleavage. We tested PLpro-ERNuc model recombinant expressed HeLa cells. demonstrated expected reticular network channels absence protease, efficient translocation nuclei PLpro-expressing (14-fold increase nucleus/cytoplasm ratio). Then, Huh7.5 infected SARS-CoV-2 virus, where it showed robust ER-to-nucleus 24 h post infection. believe that represents useful tool screening inhibitors monitoring virus spread culture.

Language: Английский

Citations

0
Natural-Target-Mimicking Translocation-Based Fluorescent Sensor for Detection of SARS-CoV-2 PLpro Protease Activity and Virus Infection in Living Cells DOI Open Access
Elena L. Sokolinskaya, Olga N. Ivanova,

Irina T. Fedyakina

et al.

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(12), P. 6635 - 6635

Published: June 17, 2024

Papain-like protease PLpro, a domain within large polyfunctional protein, nsp3, plays key roles in the life cycle of SARS-CoV-2, being responsible for first events cleavage polyprotein into individual proteins (nsp1-4) as well suppression cellular immunity. Here, we developed new genetically encoded fluorescent sensor, named PLpro-ERNuc, detection PLpro activity living cells using translocation-based readout. The sensor was designed follows. A fragment nsp3 protein used to direct on cytoplasmic surface endoplasmic reticulum (ER) membrane, thus closely mimicking natural target PLpro. part included two bright proteins-red mScarlet I and green mNeonGreen-separated by linker with site. nuclear localization signal (NLS) attached ensure accumulation mNeonGreen nucleus upon cleavage. We tested PLpro-ERNuc model recombinant expressed HeLa cells. demonstrated expected reticular network red channels absence protease, efficient translocation nuclei PLpro-expressing (14-fold increase nucleus/cytoplasm ratio). Then, Huh7.5 infected SARS-CoV-2 virus, where it showed robust ER-to-nucleus 24 h post infection. believe that represents useful tool screening inhibitors monitoring virus spread culture.

Language: Английский

Citations

0
A guanidine-based coronavirus replication inhibitor which targets the nsp15 endoribonuclease and combines a direct antiviral effect with subtle boosting of interferon DOI Creative Commons
Benjamin Van Loy, Eugènia Pujol, Kenichi Kamata

et al.

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

Published: Sept. 9, 2024

Abstract The approval of COVID-19 vaccines and two classes antiviral drugs has been crucial in addressing the global health crisis caused by SARS-CoV-2. However, to prepare for future outbreaks from drug-resistant variants novel zoonotic coronaviruses (CoVs), additional therapeutics with a distinct mechanism are needed. Here, we report guanidine-substituted diphenylurea compound that suppresses CoV replication interfering uridine-specific endoribonuclease (EndoU) activity viral non-structural protein-15 (nsp15). This compound, designated EPB-113, exhibits strong selective cell culture against human coronavirus 229E (HCoV-229E) also HCoV-229E viruses selected resistance EPB-113 carried mutations at or near catalytic residue His250 nsp15 EndoU domain. is known reduce levels dsRNA avoid induction type I interferon (IFN-I). Accordingly, mutant under were unable replicate macrophages readily outcompeted wild-type virus upon co-infection fibroblast cells. highlights critical role maintaining replication. By targeting nsp15, combines direct effect subtle boosting IFN-I. Using combination biophysical enzymatic assays recombinant proteins SARS-CoV-2, discovered reduces thermal stability hexameric while enhancing its cleavage activity. EndoU-stimulating offers an explanation why escapes becoming EndoU-deficient. Our findings suggest as strategy may require alters conformation this protein disrupts other functions related immune evasion Based on appealing pharmacological profile conclude challenging but highly relevant drug target. Author summary Despite significant progress controlling through drugs, SARS-CoV-2 infection remains serious threat vulnerable individuals, such elderly immunocompromised patients. These populations could benefit innovative treatments target multiple angles. In study, focus (EndoU), poorly understood aids host innate response assumed regulate RNA synthesis. research centers around newly identified, first-in-class inhibitor, which specifically targets nsp15. unique property suppress complementary ways via combined impact selects escape mutants carry substitutions core EndoU, leading severe defect. When incubated protein, induces structural change enhances enzyme Hence, our study introduces concept multifunctional fronts.

Language: Английский

Citations

0
Human coronaviruses: activation and antagonism of innate immune responses DOI
Nikhila S. Tanneti, Helen A. Stillwell, Susan R. Weiss

et al.

Microbiology and Molecular Biology Reviews, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 19, 2024

SUMMARY Human coronaviruses cause a range of respiratory diseases, from the common cold (HCoV-229E, HCoV-NL63, HCoV-OC43, and SARS-CoV-2) to lethal pneumonia (SARS-CoV, SARS-CoV-2, MERS-CoV). Coronavirus interactions with host innate immune antiviral responses are an important determinant disease outcome. This review compares host’s response different human coronaviruses. Host defenses discussed in this include frontline against viruses nasal epithelium, early sensing viral infection by effectors, double-stranded RNA stress-induced pathways, antagonism conferred conserved coronavirus nonstructural proteins genus-specific accessory proteins. The HCoV-229E -NL63 induce robust interferon signaling related SARS-CoV SARS-CoV-2 intermediate levels activation, MERS-CoV shuts down these pathways almost completely.

Language: Английский

Citations

0