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Record W4297399794 · doi:10.1101/2022.09.26.509485

Allosteric regulation and crystallographic fragment screening of SARS-CoV-2 NSP15 endoribonuclease

2022· preprint· en· W4297399794 on OpenAlexfundno aff
André S. Godoy, A.M. Nakamura, A. Douangamath, Yun Song, G.D. Noske, Victor O. Gawriljuk, R.S. Fernandes, Humberto D'Muniz Pereira, Ketllyn Irene Zagato de Oliveira, D. Fearon, Alexandre Dias, T. Krojer, M. Fairhead, Alisa Powell, Louise Dunnet, J. Brandão-Neto, R. Skyner, R. Chalk, F. von Delft, Dávid Bajusz, Miklós Bege, Anikó Borbás, György M. Keserű, Glaucius Oliva

Bibliographic record

VenuebioRxiv (Cold Spring Harbor Laboratory) · 2022
Typepreprint
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicRNA and protein synthesis mechanisms
Canadian institutionsnot available
FundersCentro Nacional de Pesquisa em Energia e MateriaisBiotechnology and Biological Sciences Research CouncilNemzeti Kutatási Fejlesztési és Innovációs HivatalVetenskapsrådetMinistério da Ciência, Tecnologia e InovaçãoCoordenação de Aperfeiçoamento de Pessoal de Nível SuperiorEuropean Federation of Pharmaceutical Industries and AssociationsSvenska Forskningsrådet FormasMedical Research CouncilVINNOVAEuropean CommissionInnovációs és Technológiai MinisztériumMagyar Tudományos AkadémiaFundação de Amparo à Pesquisa do Estado de São PauloMcGill UniversityDiamond Light SourceWellcome Trust
KeywordsEndoribonucleaseAllosteric regulationEnzymeComputational biologyChemistryAllosteric enzymeBiologyBiochemistryStereochemistryRNAGene

Abstract

fetched live from OpenAlex

Abstract SARS-CoV-2 is the causative agent of COVID-19. The highly conserved viral NSP15 endoribonuclease, NendoU, is a key enzyme involved in viral immune evasion, and a promising target for the development of new classes of antivirals. Yet, the broad variety of recognition sequences, complex assembly and kinetics, and lack of structural complexes hampers the development of new competitive or allosteric inhibitors for this target. Here, we performed enzymatic characterization of NendoU in its monomeric and hexameric form, showing that hexamers are allosteric enzymes with a positive cooperative index of 2. By using cryo-EM at distinct pH’s combined with X-ray crystallography and structural analysis, we demonstrate the potential for NendoU to shift between open and closed states, and assembly in larger supramolecular entities, which might serve as a mechanism of self-regulation. Further, we report results from a large fragment screening campaign against NendoU, revealing multiple new allosteric sites for the development of inhibitors.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.001
Threshold uncertainty score0.003

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0010.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.018
GPT teacher head0.230
Teacher spread0.211 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations4
Published2022
Admission routes1
Has abstractyes

Explore more

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