Discovery of 2-Amide-3-methylester Thiophenes that Target SARS-CoV-2 Mac1 and Repress Coronavirus Replication, Validating Mac1 as an Antiviral Target

Journal of Medicinal Chemistry, Journal Year: 2024, Volume and Issue: 67(8), P. 6519 - 6536

Published: April 9, 2024

The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has made it clear that further development of antiviral therapies will be needed. Here, we describe small-molecule inhibitors for SARS-CoV-2 Mac1, which counters ADP-ribosylation-mediated innate immune responses. Three high-throughput screening hits had the same 2-amide-3-methylester thiophene scaffold. We studied compound binding mode using X-ray crystallography, allowing us to design analogues. Compound 27 (MDOLL-0229) an IC50 2.1 μM and was selective CoV Mac1 proteins after profiling activity against a panel viral human proteins. improved potency allowed testing its effect on replication, indeed, inhibited replication both murine hepatitis (MHV) prototypes SARS-CoV-2. Sequencing drug-resistant MHV identified mutations in demonstrating specificity 27. is first Mac1-targeted small molecule demonstrated inhibit cell model.

Language: Английский

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Recurrent Loss of Macrodomain Activity in Host Immunity and Viral Proteins

Pathogens, Journal Year: 2023, Volume and Issue: 12(5), P. 674 - 674

Published: May 3, 2023

Protein post-translational modifications (PTMs) are an important battleground in the evolutionary arms races that waged between host innate immune system and viruses. One such PTM, ADP-ribosylation, has recently emerged as mediator of antiviral immunity. Important for host–virus conflict over this PTM is addition ADP-ribose by PARP proteins removal macrodomain-containing proteins. Interestingly, several proteins, known macroPARPs, contain macrodomains well a domain, these both response evolving under very strong positive (diversifying) selection. In addition, viruses, including alphaviruses coronaviruses, encode one or more macrodomains. Despite presence conserved macrodomain fold, enzymatic activity many not been characterized. Here, we perform functional analyses to characterize macroPARP viral We trace history macroPARPs metazoans show PARP9 PARP14 single active macrodomain, whereas PARP15 contains none. also reveal independent losses within mammalian PARP14, bat, ungulate, carnivore lineages. Similar coronaviruses up three macrodomains, with only first displaying catalytic activity. Intriguingly, recurrent loss alphavirus group insect-specific two human-infecting Together, our data unexpected turnover

Language: Английский

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Anti-viral defense by an ADP-ribosyltransferase that targets mRNA to block translation

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

Published: Feb. 24, 2024

Abstract Host-pathogen conflicts are crucibles of molecular innovation. Selection for immunity to pathogens has driven the evolution sophisticated mechanisms throughout biology, including in bacteria that must evade their viral predators known as bacteriophages. Here, we characterize a widely distributed anti-phage defense system, CmdTAC, provides robust against infection by T-even family phages. Our results support model which CmdC detects sensing capsid proteins, ultimately leading activation toxic ADP-ribosyltransferase effector protein, CmdT. We show newly synthesized protein triggers dissociation chaperone from CmdTAC complex, destabilization and degradation antitoxin CmdA, with consequent liberation CmdT ADP-ribosyltransferase. Strikingly, does not target DNA, or structured RNA, targets other ADP-ribosyltransferases. Instead, modifies N6 position adenine GA dinucleotides within single-stranded RNAs arrest mRNA translation inhibition replication. work reveals new mechanism anti-viral previously unknown but broadly class ADP-ribosyltransferases mRNA.

Language: Английский

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Mono-ADP-ribosylation by PARP10 inhibits Chikungunya virus nsP2 proteolytic activity and viral replication

Cellular and Molecular Life Sciences, Journal Year: 2023, Volume and Issue: 80(3)

Published: Feb. 25, 2023

Replication of viruses requires interaction with host cell factors and repression innate immunity. Recent findings suggest that a subset intracellular mono-ADP-ribosylating PARPs, which are induced by type I interferons, possess antiviral activity. Moreover, certain RNA viruses, including Chikungunya virus (CHIKV), encode mono-ADP-ribosylhydrolases. Together, this suggests role for mono-ADP-ribosylation (MARylation) in host-virus conflicts, but the relevant substrates have not been identified. We addressed PARP restricts CHIKV replication identified PARP10 PARP12. For PARP10, restriction was dependent on catalytic processing non-structural polyprotein nsP1-4 protease located nsP2 assembly four individual nsP1-nsP4 into functional complex. PARP12 inhibited production nsP3, indicating defect processing. The nsP3 protein encodes macrodomain de-MARylation activity, is essential replication. In support MARylation affecting processing, defective replicons revealed reduced nsP3. hypothesized regulates proteolytic function nsP2. Indeed, we found MARylated and, as consequence, its activity inhibited. NsP3-dependent reactivated protease. Hence, propose PARP10-mediated prevents consequently our provide mechanistic explanation viral MAR hydrolase

Language: Английский

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Emerging Role of Nicotinamide Riboside in Health and Diseases

Nutrients, Journal Year: 2022, Volume and Issue: 14(19), P. 3889 - 3889

Published: Sept. 20, 2022

Among all the NAD+ precursors, nicotinamide riboside (NR) has gained most attention as a potent NAD+-enhancement agent. This recently discovered vitamin, B3, demonstrated excellent safety and efficacy profiles is orally bioavailable in humans. Boosting intracellular concentrations using NR been shown to provide protective effects against broad spectrum of pathological conditions, such neurodegenerative diseases, diabetes, hearing loss. In this review, an integrated overview research will be presented. The role plays biosynthetic pathway introduced, followed by discussion on synthesis chemical enzymatic approaches. NR’s regulating normal physiology pathophysiology also presented, focusing studies published last five years.

Language: Английский

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Zinc-finger PARP proteins ADP-ribosylate alphaviral proteins and are required for interferon-γ–mediated antiviral immunity

Science Advances, Journal Year: 2025, Volume and Issue: 11(5)

Published: Jan. 31, 2025

Viral manipulation of posttranslational modifications (PTMs) is critical to enable control over host defenses. Evidence suggests that one such PTM, adenosine 5′-diphosphate (ADP)–ribosylation, important for viral replication, but the and components involved are poorly understood. Here, we demonstrate several human poly(ADP-ribose) polymerase (PARP) proteins, including zinc-finger domain containing PARP7 (TiPARP) PARP12, directly ADP-ribosylate alphaviral nonstructural proteins (nsPs), nsP3 nsP4. These same PARP inhibit alphavirus replication in a manner can be antagonized by ADP-ribosylhydrolase activity virally encoded macrodomain. Last, find knockdown any three CCCH PARPs, PARP7, or enzymatically inactive PARP13 (ZAP/ZC3HAV1), attenuates antiviral effects interferon-γ on replication. Combined with evolutionary analyses, these data suggest PARPs share an ancestral function macrodomains, indicative ongoing conflict between ADP-ribosylation viruses.

Language: Английский

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Derivatives of MOPS: promising scaffolds for SARS coronaviruses Macro domain‐targeted inhibition

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

Published: March 11, 2025

The severe acute respiratory syndrome coronavirus (SARS‐CoV/CoV‐2) genome encodes 16 non‐structural proteins (nsps), which coordinate cell remodeling, virus replication and participate in viral evasion. Notably, nsp3 contains a protein module termed Macro domain, carries IFN antagonist activity that interferes with host innate immunity response. This domain is able to bind hydrolyze ADP‐ribose derivatives. correlated escape thus makes domains valuable therapeutic target. In the present paper, we report SARS‐CoV structure complex MOPS molecule. Based on our structural data, molecular docking was performed set of analogs binding pocket. We an ELISA‐based assay select hits based inhibition recombinant SARS‐CoV/CoV‐2 domain‐ADP‐ribose formation. Among tested analogs, MOPSO CAPSO are more efficient inhibiting ADP‐ribose‐binding. Structural analysis these molecules pocket reveals potential interactions amino acid residues involved coordination ADP‐ribose. Overall, findings suggest bear be used as scaffold for design domain‐specific inhibitors.

Language: Английский

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Host–Pathogen Interaction Interface: Promising Candidate Targets for Vaccine-Induced Protective and Memory Immune Responses

Vaccines, Journal Year: 2025, Volume and Issue: 13(4), P. 418 - 418

Published: April 16, 2025

Vaccine formulations are a successful strategy against pathogen transmission because vaccine candidates induce effective and long-lasting memory immune responses (B CD4+ T cells) at systemic mucosal sites. Extracellular vesicles of lipoproteins, bioactive compounds from plants invertebrates (sponges) encapsulated in liposomes, glycoproteins can target these The developed mimic microbial pathogens way that successfully links the innate adaptive responses. In addition, vaccines plus adjuvants promote maintain an inflammatory response. this review, we aimed to identify host–pathogen interface as rich source candidate targets for vaccine-induced protective

Language: Английский

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Monitoring nucleolar-nucleoplasmic protein shuttling in living cells by high-content microscopy and automated image analysis

Nucleic Acids Research, Journal Year: 2024, Volume and Issue: 52(15), P. e72 - e72

Published: June 26, 2024

Abstract The nucleolus has core functions in ribosome biosynthesis, but also acts as a regulatory hub plethora of non-canonical processes, including cellular stress. Upon DNA damage, several repair factors shuttle between the and nucleoplasm. Yet, molecular mechanisms underlying such spatio-temporal protein dynamics remain to be deciphered. Here, we present novel imaging platform investigate nucleolar-nucleoplasmic shuttling living cells. For image acquisition, used commercially available automated fluorescence microscope for analysis, developed KNIME workflow with implementation machine learning-based tools. We validated method different nucleolar proteins, i.e., PARP1, TARG1 APE1, by monitoring their upon oxidative As paradigm, analyzed PARP1 H2O2 treatment combination range pharmacological inhibitors reporter cell line. These experiments revealed that inhibition SIRT7 results loss localization. Finally, unraveled specific differences after co-treatment clinical PARP inhibitors. Collectively, this work delineates highly sensitive versatile bioimaging swift cells, which can employed screening in-depth mechanistic analyses.

Language: Английский

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PARP enzymes and mono-ADP-ribosylation: advancing the connection from interferon-signalling to cancer biology

Expert Reviews in Molecular Medicine, Journal Year: 2024, Volume and Issue: 26

Published: Jan. 1, 2024

Abstract ADP-ribosyltransferases of the PARP family encompass a group enzymes with variegated regulatory functions in cells, ranging from DNA damage repair to control cell-cycle progression and immune response. Over years, this knowledge has led use PARP1/2 inhibitors as mainstay pharmaceutical strategies for treatment ovarian, pancreatic, prostate breast cancers, holding mutations genes encoding proteins involved mechanisms (synthetic lethality). Meanwhile, last decade witnessed significant progress comprehending cellular pathways regulated by mono-ADP-ribosylation, huge effort development novel selective compounds inhibit those PARPs endowed mono-ADP-ribosylation activity. This review focuses on achieved cancer field, delving into most recent findings regarding role subset – interferon-stimulated progression.

Language: Английский

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