Structure and function of yeast Lso2 and human CCDC124 bound to hibernating ribosomes

Wells, Jennifer N.; Buschauer, Robert; Mackens-Kiani, Timur; Best, Katharina; Kratzat, Hanna; Berninghausen, Otto; Becker, Thomas; Gilbert, Wendy V.; Cheng, Jingdong; Beckmann, Roland

doi:10.1101/2020.02.12.944066

Cited by 14 publications

(19 citation statements)

References 69 publications

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“…S4, A to E) contained the protein coiled-coil domain containing short open reading frame 124 (CCDC124), a homolog of the ribosome protection and translation recovery factor Lso2 in Saccharomyces cerevisiae (29). A similar complex of inactive 80S ribosomes bound to CCDC124 was recently described (30). In contrast to the known hibernation complex, however, a subpopulation of the Nsp1-bound complex contained in addition the ribosome recycling factor and ABCtype ATPase ABCE1 (31-33) together with class I translation termination factor eRF1 in an unusual conformation ( fig.…”

Section: In Vitro Reconstituted and Native Nsp1-40s And -80s Ribosomementioning

confidence: 99%

Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2

Thoms¹,

Buschauer²,

Ameismeier³

et al. 2020

Preprint

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160

232

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SARS-CoV-2 is the causative agent of the current COVID-19 pandemic. A major virulence factor of SARS-CoVs is the nonstructural protein 1 (Nsp1) which suppresses host gene expression by ribosome association via an unknown mechanism. Here, we show that Nsp1 from SARS-CoV-2 binds to 40S and 80S ribosomes, resulting in shutdown of capped mRNA translation both in vitro and in cells. Structural analysis by cryo-electron microscopy (cryo-EM) of in vitro reconstituted Nsp1-40S and of native human Nsp1-ribosome complexes revealed that the Nsp1 C-terminus binds to and obstructs the mRNA entry tunnel. Thereby, Nsp1 effectively blocks RIG-Idependent innate immune responses that would otherwise facilitate clearance of the infection. Thus, the structural characterization of the inhibitory mechanism of Nsp1 may aid structure-based drug design against SARS-CoV-2.Coronaviruses (CoVs) are enveloped, single-stranded viruses

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Section: In Vitro Reconstituted and Native Nsp1-40s And -80s Ribosomementioning

confidence: 99%

Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2

Thoms¹,

Buschauer²,

Ameismeier³

et al. 2020

Preprint

Self Cite

160

232

View full text Add to dashboard Cite

show abstract

“…S4, A to E) contained the protein coiled-coil domain containing short open reading frame 124 (CCDC124), a homolog of the ribosome protection and translation recovery factor Lso2 in Saccharomyces cerevisiae (29). A similar complex of inactive 80S ribosomes bound to CCDC124 was recently described (30). In addition to the known hibernation complex, a subpopulation of the CCDC124-bound 80S contained also the ribosome recycling factor and ABC-type ATPase ABCE1 (31-33) and the class I translation termination factor eRF1 in an unusual conformation ( fig.…”

mentioning

confidence: 99%

Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2

Thoms¹,

Buschauer²,

Ameismeier³

et al. 2020

Science

Self Cite

744

846

View full text Add to dashboard Cite

SARS-CoV-2 is the causative agent of the current COVID-19 pandemic. A major virulence factor of SARS-CoVs is the nonstructural protein 1 (Nsp1) which suppresses host gene expression by ribosome association. Here, we show that Nsp1 from SARS-CoV-2 binds to the 40S ribosomal subunit, resulting in shutdown of mRNA translation both in vitro and in cells. Structural analysis by cryo-electron microscopy (cryo-EM) of in vitro reconstituted Nsp1-40S and various native Nsp1-40S and -80S complexes revealed that the Nsp1 C terminus binds to and obstructs the mRNA entry tunnel. Thereby, Nsp1 effectively blocks RIG-I-dependent innate immune responses that would otherwise facilitate clearance of the infection. Thus, the structural characterization of the inhibitory mechanism of Nsp1 may aid structure-based drug design against SARS-CoV-2.

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“…Lee et al, 2014), its dissociation from ribosomes or even its degradation during infection could lead to activation of IRES function. In this context it would be of interest to determine the potential influence of other 80S-binding proteins such as IFRD2 and CCDC124 (Brown et al, 2018;Wells et al, 2020) on the activity of HalV and canonical IGR IRESs.…”

Section: Discussionmentioning

confidence: 99%

The Halastavi árva virus intergenic region IRES promotes translation by the simplest possible initiation mechanism

Abaeva

Vicens

Bochler

et al. 2020

Preprint

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Dicistrovirus intergenic region internal ribosomal entry sites (IGR IRES) do not require initiator tRNA, an AUG codon or initiation factors, and jumpstart translation from the middle of the elongation cycle via formation of IRES/80S complexes resembling the pre-translocation state. eEF2 then translocates the [codon-anticodon]-mimicking pseudoknot I (PKI) from ribosomal A to P sites, bringing the first sense codon into the decoding center. Halastavi árva virus (HalV) contains an IGR that is related to previously described IGR IRESs, but lacks domain 2, which enables these IRESs to bind to individual 40S ribosomal subunits. By employing in vitro reconstitution and cryo-electron microscopy, we now report that the HalV IGR IRES functions by the simplest initiation mechanism that involves binding to 80S ribosomes such that PKI is placed in the P site, so that the A site contains the first codon that is directly accessible for decoding without prior eEF2-mediated translocation of PKI.

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Structure and function of yeast Lso2 and human CCDC124 bound to hibernating ribosomes

Cited by 14 publications

References 69 publications

Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2

Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2

Structural basis for translational shutdown and immune evasion by the Nsp1 protein of SARS-CoV-2

The Halastavi árva virus intergenic region IRES promotes translation by the simplest possible initiation mechanism

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