Human coronavirus NL63 nsp1 induces degradation of RNA polymerase II to inhibit host protein synthesis DOI Creative Commons

Kala Hardy,

Michael M. Lutz, Toru Takimoto

et al.

PLoS Pathogens, Journal Year: 2024, Volume and Issue: 20(6), P. e1012329 - e1012329

Published: June 20, 2024

Coronavirus (CoV) nonstructural protein 1 (nsp1) is considered a pathogenic factor due to its ability inhibit host antiviral responses by inducing general shutoff of synthesis. Nsp1 expressed α- and β-CoVs, but functions strategies induce are not fully elucidated. We compared the nsp1s from two β-CoVs (SARS-CoV SARS-CoV-2) α-CoVs (NL63 229E) found that NL63 nsp1 has strongest activity. Unlike SARS-CoV nsp1s, which bind 40S ribosomes block translation cellular mRNA, did mRNAs transfected into cells. Instead, localized nucleus specifically inhibited transcription genes under an RNA polymerase II (RNAPII) promoter. Further analysis revealed induces degradation largest subunit RNAPII, Rpb1. This was detected regardless phosphorylation state Rpb1 blocked proteasome inhibitor MG132. also ubiquitinated in NL63-infected cells, inhibition ubiquitination ubiquitin activating enzyme (TAK243) prevented virus-infected These data reveal unrecognized strategy human α-CoV NL63: targeting prevent expression. Our study indicates viruses within same family can use completely distinct mechanisms regulate responses.

Language: Английский

Substrate recognition principles for the PP2A-B55 protein phosphatase DOI Creative Commons
Thomas Kruse,

Dimitriya H. Garvanska,

Julia K. Varga

et al.

Science Advances, Journal Year: 2024, Volume and Issue: 10(40)

Published: Oct. 2, 2024

The PP2A-B55 phosphatase regulates a plethora of signaling pathways throughout eukaryotes. How selects its substrates presents severe knowledge gap. By integrating AlphaFold modeling with comprehensive high-resolution mutational scanning, we show that α helices in bind B55 through an evolutionary conserved mechanism. Despite large diversity sequence and composition, these share key amino acid determinants engage discrete hydrophobic electrostatic patches. Using deep learning protein design, generate specific potent competitive peptide inhibitor substrate interactions. With this inhibitor, uncover the nuclear exosome targeting (NEXT) complex by binding to α-helical recruitment module RNA 7 (RBM7), component NEXT complex. Collectively, our findings provide framework for understanding interrogation function health disease.

Language: Английский

Citations

7
The transcriptional and translational landscape of HCoV-OC43 infection DOI Creative Commons
Stefan Bresson,

Emanuela Sani,

Alicja Armatowska

et al.

PLoS Pathogens, Journal Year: 2025, Volume and Issue: 21(1), P. e1012831 - e1012831

Published: Jan. 27, 2025

The coronavirus HCoV-OC43 circulates continuously in the human population and is a frequent cause of common cold. Here, we generated high-resolution atlas transcriptional translational landscape OC43 during time course following infection lung fibroblasts. Using ribosome profiling, quantified relative expression canonical open reading frames (ORFs) identified previously unannotated ORFs. These included several potential short upstream ORFs putative ORF nested inside M gene. In parallel, analyzed cellular response to infection. Endoplasmic reticulum (ER) stress genes were transcriptionally translationally induced beginning 12 18 hours post infection, respectively. By contrast, conventional antiviral mostly remained quiescent. At same points, observed accumulation increased translation noncoding transcripts normally targeted by nonsense mediated decay (NMD), suggesting NMD suppressed This work provides resources for deeper understanding gene responses

Language: Английский

Citations

0
The impact of Coronavirus Nsp1 on host mRNA degradation is independent of its role in translation inhibition DOI Creative Commons

Emilie Bäumlin,

Dominic Andenmatten,

Jonas Luginbühl

et al.

Cell Reports, Journal Year: 2025, Volume and Issue: 44(4), P. 115488 - 115488

Published: March 29, 2025

Language: Английский

Citations

0
SARS-CoV-2 targets ribosomal RNA biogenesis DOI Creative Commons
V. Talya Yerlici, Audrey Astori,

Nevraj S. Kejiou

et al.

Cell Reports, Journal Year: 2024, Volume and Issue: 43(3), P. 113891 - 113891

Published: Feb. 29, 2024

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) hinders host gene expression, curbing defenses and licensing viral protein synthesis virulence. During SARS-CoV-2 infection, the virulence factor non-structural 1 (Nsp1) targets mRNA entry channel of mature cytoplasmic ribosomes, limiting translation. We show that Nsp1 also restrains translation by targeting nucleolar ribosome biogenesis. infection disrupts 18S 28S ribosomal RNA (rRNA) processing. Expression recapitulates processing defects. abrogates rRNA production without altering expression critical factors or organization. Instead, localizes to nucleolus, interacting with precursor-rRNA hindering its maturation separately from protein's role in restricting ribosomes. Thus, limits biogenesis These findings revise our understanding how controls human synthesis, suggesting efforts counter Nsp1's effect on should consider impact manufacturing

Language: Английский

Citations

3
The 5′-terminal stem–loop RNA element of SARS-CoV-2 features highly dynamic structural elements that are sensitive to differences in cellular pH DOI Creative Commons
Sabrina Toews, Anna Wacker, Edgar M. Faison

et al.

Nucleic Acids Research, Journal Year: 2024, Volume and Issue: 52(13), P. 7971 - 7986

Published: June 6, 2024

Abstract We present the nuclear magnetic resonance spectroscopy (NMR) solution structure of 5′-terminal stem loop 5_SL1 (SL1) SARS-CoV-2 genome. SL1 contains two A-form helical elements and regions with non-canonical structure, namely an apical pyrimidine-rich asymmetric internal one nucleotides at 5′- 3′-terminal part sequence, respectively. The conformational ensemble representing averaged was validated using NMR residual dipolar coupling (RDC) small-angle X-ray scattering (SAXS) data. show that is major binding site for fragments low molecular weight. This can be stabilized by A12–C28 interaction promotes transient formation A+•C base pair. As a consequence, pKa adenosine A12 shifted to 5.8, compared 3.63 free adenosine. Furthermore, applying recently developed pH-differential mutational profiling (PD-MaP) approach, we not only recapitulated our findings but also unveiled multiple sites potentially sensitive pH across 5′-UTR SARS-CoV-2.

Language: Английский

Citations

3
Alpha- and betacoronavirus cis-acting RNA elements DOI Creative Commons
Ramakanth Madhugiri, Nguyễn Hoàng Việt, Heiko Slanina

et al.

Current Opinion in Microbiology, Journal Year: 2024, Volume and Issue: 79, P. 102483 - 102483

Published: May 8, 2024

Coronaviruses have exceptionally large RNA genomes and employ multiprotein replication/transcription complexes to orchestrate specific steps of viral genome replication expression. Most these processes involve cis-acting elements that are engaged in vital RNA-RNA and/or RNA-protein interactions. Over the past years, a number studies provided interesting new insight into structures and, lesser extent, functions for representative coronaviruses, there is evidence suggest (a majority of) conserved across genetically divergent coronavirus genera. It becoming increasingly clear at least some do not function isolation but operate through complex highly dynamic This article reviews structural functional aspects alpha- betacoronavirus 5'- 3'-terminal regions, focusing on their critical roles synthesis gene

Language: Английский

Citations

1
Uncoupling the functional roles of Coronavirus Nsp1 DOI Creative Commons

Emilie Bäumlin,

Dominic Andenmatten,

Jonas Luginbühl

et al.

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

Published: Aug. 5, 2024

Summary When host cells are infected with coronaviruses, the first viral protein produced is Nsp1. This inhibits synthesis and induces mRNA degradation to enhance proliferation. Despite its critical role, mechanism by which Nsp1 mediates cellular remains unclear. In this study, we use cell-free translation address how stability regulated We reveal that SARS-CoV-2 binding ribosome enough trigger independently of collisions or active translation. MERS-CoV without triggering degradation, highlighting mechanistic differences between two counterparts. mRNAs appear co-evolve, rendering immune Nsp1-mediated in SARS-CoV-2, Bat-Hp viruses. By providing new insights into mode action Nsp1, our study helps understand biology better find strategies for therapeutic targeting against coronaviral infections. Significance Cell-free assays allow decoupling inhibition from RNA degradation. interaction crucial but not required. while 5’UTR-specific protection indicates a co-evolutionary adaptation features

Language: Английский

Citations

1
The emerging role of SARS-CoV-2 non-structural protein 1 (nsp1) in epigenetic regulation of host gene expression DOI Creative Commons
Konstantin I. Ivanov, Haibin Yang, Ruixue Sun

et al.

FEMS Microbiology Reviews, Journal Year: 2024, Volume and Issue: 48(5)

Published: Sept. 1, 2024

Infection with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes widespread changes in epigenetic modifications and chromatin architecture host cell. Recent evidence suggests that SARS-CoV-2 nonstructural protein 1 (nsp1) plays an important role driving these changes. Previously thought to be primarily involved translation shutoff cellular mRNA degradation, nsp1 has now been shown a truly multifunctional affects gene expression at multiple levels. The functions of are surprisingly diverse include not only downregulation stability, but also inhibition export from nucleus, suppression immune signaling, and, most recently, regulation expression. In this review, we first summarize current knowledge on SARS-CoV-2-induced structure. We then focus reprogramming, particular emphasis silencing immune-related genes. Finally, discuss potential molecular mechanisms underlying based interactome studies.

Language: Английский

Citations

1
Human coronavirus NL63 nsp1 induces degradation of RNA polymerase II to inhibit host protein synthesis DOI Creative Commons

Kala Hardy,

Michael M. Lutz, Toru Takimoto

et al.

PLoS Pathogens, Journal Year: 2024, Volume and Issue: 20(6), P. e1012329 - e1012329

Published: June 20, 2024

Coronavirus (CoV) nonstructural protein 1 (nsp1) is considered a pathogenic factor due to its ability inhibit host antiviral responses by inducing general shutoff of synthesis. Nsp1 expressed α- and β-CoVs, but functions strategies induce are not fully elucidated. We compared the nsp1s from two β-CoVs (SARS-CoV SARS-CoV-2) α-CoVs (NL63 229E) found that NL63 nsp1 has strongest activity. Unlike SARS-CoV nsp1s, which bind 40S ribosomes block translation cellular mRNA, did mRNAs transfected into cells. Instead, localized nucleus specifically inhibited transcription genes under an RNA polymerase II (RNAPII) promoter. Further analysis revealed induces degradation largest subunit RNAPII, Rpb1. This was detected regardless phosphorylation state Rpb1 blocked proteasome inhibitor MG132. also ubiquitinated in NL63-infected cells, inhibition ubiquitination ubiquitin activating enzyme (TAK243) prevented virus-infected These data reveal unrecognized strategy human α-CoV NL63: targeting prevent expression. Our study indicates viruses within same family can use completely distinct mechanisms regulate responses.

Language: Английский

Citations

0