{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T00:09:55Z","timestamp":1788480595253,"version":"build-2803163510"},"reference-count":67,"publisher":"Oxford University Press (OUP)","issue":"10","license":[{"start":{"date-parts":[[2023,4,28]],"date-time":"2023-04-28T00:00:00Z","timestamp":1682640000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["88887.516153\/2020-00"],"award-info":[{"award-number":["88887.516153\/2020-00"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2013\/07600-3"],"award-info":[{"award-number":["2013\/07600-3"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2015\/16811-3"],"award-info":[{"award-number":["2015\/16811-3"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2016\/19712-9"],"award-info":[{"award-number":["2016\/19712-9"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"name":"EU\/EFPIA\/OICR\/McGill\/KTH\/Diamond Innovative Medicines Initiative 2","award":["EUbOPEN 875510"],"award-info":[{"award-number":["EUbOPEN 875510"]}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["U19AI171399"],"award-info":[{"award-number":["U19AI171399"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011019","name":"National Research Development and Innovation Office","doi-asserted-by":"publisher","award":["SNN 135335"],"award-info":[{"award-number":["SNN 135335"]}],"id":[{"id":"10.13039\/501100011019","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003825","name":"Hungarian Academy of Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003825","id-type":"DOI","asserted-by":"publisher"}]},{"name":"\u00daNKP-22-5 New National Excellence Program of the Ministry for Innovation and Technology"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,6,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). The NSP15 endoribonuclease enzyme, known as NendoU, is highly conserved and plays a critical role in the ability of the virus to evade the immune system. NendoU is a promising target for the development of new antiviral drugs. However, the complexity of the enzyme's structure and kinetics, along with the broad range of recognition sequences and lack of structural complexes, hampers the development of inhibitors. Here, we performed enzymatic characterization of NendoU in its monomeric and hexameric form, showing that hexamers are allosteric enzymes with a positive cooperative index, and with no influence of manganese on enzymatic activity. Through combining cryo-electron microscopy at different pHs, X-ray crystallography and biochemical and structural analysis, we showed that NendoU can shift between open and closed forms, which probably correspond to active and inactive states, respectively. We also explored the possibility of NendoU assembling into larger supramolecular structures and proposed a mechanism for allosteric regulation. In addition, we conducted a large fragment screening campaign against NendoU and identified several new allosteric sites that could be targeted for the development of new inhibitors. Overall, our findings provide insights into the complex structure and function of NendoU and offer new opportunities for the development of inhibitors.<\/jats:p>","DOI":"10.1093\/nar\/gkad314","type":"journal-article","created":{"date-parts":[[2023,4,28]],"date-time":"2023-04-28T09:39:51Z","timestamp":1682674791000},"page":"5255-5270","source":"Crossref","is-referenced-by-count":26,"title":["Allosteric regulation and crystallographic fragment screening of SARS-CoV-2 NSP15 endoribonuclease"],"prefix":"10.1093","volume":"51","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0613-9164","authenticated-orcid":false,"given":"Andre Schutzer","family":"Godoy","sequence":"first","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. Joao Dagnone, 1100 - Jardim Santa Angelina, Sao Carlos , 13563-120, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aline Minalli","family":"Nakamura","sequence":"additional","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. Joao Dagnone, 1100 - Jardim Santa Angelina, Sao Carlos , 13563-120, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alice","family":"Douangamath","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun","family":"Song","sequence":"additional","affiliation":[{"name":"Electron Bio-imaging Centre, Diamond Light Source Ltd. , Harwell Science and Innovation Campus, Didcot OX11 0QX, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela Dias","family":"Noske","sequence":"additional","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. Joao Dagnone, 1100 - Jardim Santa Angelina, Sao Carlos , 13563-120, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor Oliveira","family":"Gawriljuk","sequence":"additional","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. Joao Dagnone, 1100 - Jardim Santa Angelina, Sao Carlos , 13563-120, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafaela Sachetto","family":"Fernandes","sequence":"additional","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. Joao Dagnone, 1100 - Jardim Santa Angelina, Sao Carlos , 13563-120, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Humberto\u00a0D\u00a0Muniz","family":"Pereira","sequence":"additional","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. Joao Dagnone, 1100 - Jardim Santa Angelina, Sao Carlos , 13563-120, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ketllyn\u00a0Irene\u00a0Zagato","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. Joao Dagnone, 1100 - Jardim Santa Angelina, Sao Carlos , 13563-120, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daren","family":"Fearon","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Dias","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tobias","family":"Krojer","sequence":"additional","affiliation":[{"name":"BioMAX, MAX IV Laboratory , Fotongatan 2, Lund 224 84, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Fairhead","sequence":"additional","affiliation":[{"name":"Centre for Medicines Discovery, Oxford University , Oxford OX1 3QU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alisa","family":"Powell","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louise","family":"Dunnet","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose","family":"Brandao-Neto","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachael","family":"Skyner","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rod","family":"Chalk","sequence":"additional","affiliation":[{"name":"Centre for Medicines Discovery, Oxford University , Oxford OX1 3QU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4277-9481","authenticated-orcid":false,"given":"D\u00e1vid","family":"Bajusz","sequence":"additional","affiliation":[{"name":"Medicinal Chemistry Research Group, Research Centre for Natural Sciences , Magyar tud\u00f3sok krt. 2, 1117 Budapest , Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikl\u00f3s","family":"Bege","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, University of Debrecen , Egyetem t\u00e9r 1, 4032 Debrecen , Hungary"},{"name":"MTA-DE Molecular Recognition and Interaction Research Group, University of Debrecen , Egyetem t\u00e9r 1, 4032 Debrecen , Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anik\u00f3","family":"Borb\u00e1s","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Chemistry, University of Debrecen , Egyetem t\u00e9r 1, 4032 Debrecen , Hungary"},{"name":"National Laboratory of Virology, University of P\u00e9cs , Ifj\u00fas\u00e1g \u00fatja 20, H-7624 P\u00e9cs, Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gy\u00f6rgy Mikl\u00f3s","family":"Keser\u0171","sequence":"additional","affiliation":[{"name":"Medicinal Chemistry Research Group, Research Centre for Natural Sciences , Magyar tud\u00f3sok krt. 2, 1117 Budapest , Hungary"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0378-0017","authenticated-orcid":false,"given":"Frank","family":"von\u00a0Delft","sequence":"additional","affiliation":[{"name":"Diamond Light Source Ltd, Harwell Science and Innovation Campus , Didcot OX11 0QX, UK"},{"name":"Research Complex at Harwell, Harwell Science and Innovation Campus , Didcot OX11 0FA, UK"},{"name":"Centre for Medicines Discovery, Oxford University , Oxford OX1 3QU, UK"},{"name":"Department of Biochemistry, University of Johannesburg , PO Box 524, Auckland Park 2006 , South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Glaucius","family":"Oliva","sequence":"additional","affiliation":[{"name":"Sao Carlos Institute of Physics, University of Sao Paulo , Av. 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Agents Chemother."},{"key":"2023060902073677700_B4","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/S0140-6736(20)30251-8","article-title":"Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding","volume":"395","author":"Lu","year":"2020","journal-title":"Lancet North Am. Ed."},{"key":"2023060902073677700_B5","doi-asserted-by":"crossref","first-page":"6558","DOI":"10.1111\/jcmm.15364","article-title":"Scientific research progress of COVID-19\/SARS-CoV-2 in the first five months","volume":"24","author":"Li","year":"2020","journal-title":"J. Cell. Mol. Med."},{"key":"2023060902073677700_B6","doi-asserted-by":"crossref","first-page":"1306","DOI":"10.1126\/science.abf3546","article-title":"Structural basis of ribosomal frameshifting during translation of the SARS-CoV-2 RNA genome","volume":"372","author":"Bhatt","year":"2021","journal-title":"Science"},{"key":"2023060902073677700_B7","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.antiviral.2013.11.006","article-title":"SARS-CoV ORF1b-encoded nonstructural proteins 12\u201316: replicative enzymes as antiviral targets","volume":"101","author":"Subissi","year":"2014","journal-title":"Antiviral Res."},{"key":"2023060902073677700_B8","doi-asserted-by":"crossref","first-page":"1596","DOI":"10.1002\/pro.3873","article-title":"Crystal structure of Nsp15 endoribonuclease NendoU from SARS-CoV-2","volume":"29","author":"Kim","year":"2020","journal-title":"Protein Sci."},{"key":"2023060902073677700_B9","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1016\/S0022-2836(03)00865-9","article-title":"Unique and conserved features of genome and proteome of SARS-coronavirus, an early split-off from the coronavirus group 2 lineage","volume":"331","author":"Snijder","year":"2003","journal-title":"J. Mol. Biol."},{"key":"2023060902073677700_B10","doi-asserted-by":"crossref","first-page":"3655","DOI":"10.1074\/jbc.M708375200","article-title":"Structural and functional analyses of the severe acute respiratory syndrome coronavirus endoribonuclease Nsp15","volume":"283","author":"Bhardwaj","year":"2008","journal-title":"J. Biol. Chem."},{"key":"2023060902073677700_B11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41467-020-20608-z","article-title":"Cryo-EM structures of the SARS-CoV-2 endoribonuclease Nsp15 reveal insight into nuclease specificity and dynamics","volume":"12","author":"Pillon","year":"2021","journal-title":"Nat. Commun."},{"key":"2023060902073677700_B12","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1016\/j.jmb.2005.09.007","article-title":"Mutational analysis of the SARS virus Nsp15 endoribonuclease: identification of residues affecting hexamer formation","volume":"353","author":"Guarino","year":"2005","journal-title":"J. Mol. 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Commun."},{"key":"2023060902073677700_B16","doi-asserted-by":"crossref","first-page":"E4251","DOI":"10.1073\/pnas.1618310114","article-title":"Coronavirus nonstructural protein 15 mediates evasion of dsRNA sensors and limits apoptosis in macrophages","volume":"114","author":"Deng","year":"2017","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"2023060902073677700_B17","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.virol.2017.12.024","article-title":"An \u2018old\u2019 protein with a new story: coronavirus endoribonuclease is important for evading host antiviral defenses","volume":"517","author":"Deng","year":"2018","journal-title":"Virology"},{"key":"2023060902073677700_B18","doi-asserted-by":"crossref","first-page":"1418","DOI":"10.1080\/22221751.2020.1780953","article-title":"SARS-CoV-2 nsp13, nsp14, nsp15 and orf6 function as potent interferon antagonists","volume":"9","author":"Yuen","year":"2020","journal-title":"Emerg. 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