Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages

RNase2 is the member of the RNaseA family most abundant in macrophages. Here, we knocked out RNase2 in THP-1 cells and analysed the response to Respiratory Syncytial Virus (RSV). RSV induced RNase2 expression, which significantly enhanced cell survival. Next, by cP-RNAseq sequencing, which amplifies...

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Autores: Lu, Lu, Li, Jiarui, Wei, Ranlei, Guidi, Irene, Cozzuto, Luca, Ponomarenko, Julia, Prats-Ejarque, Guillem, Boix, Ester
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2022
País:España
Institución:Universitat Pompeu Fabra
Repositorio:Repositorio Digital de la UPF
OAI Identifier:oai:repositori.upf.edu:10230/53264
Acceso en línea:http://hdl.handle.net/10230/53264
http://dx.doi.org/10.1007/s00018-022-04229-x
Access Level:acceso abierto
Palabra clave:Genètica
Virus
Virus sincicial respiratori
Ribonucleases
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spelling Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophagesLu, LuLi, JiaruiWei, RanleiGuidi, IreneCozzuto, LucaPonomarenko, JuliaPrats-Ejarque, GuillemBoix, EsterGenèticaVirusVirus sincicial respiratoriRibonucleasesRNase2 is the member of the RNaseA family most abundant in macrophages. Here, we knocked out RNase2 in THP-1 cells and analysed the response to Respiratory Syncytial Virus (RSV). RSV induced RNase2 expression, which significantly enhanced cell survival. Next, by cP-RNAseq sequencing, which amplifies the cyclic-phosphate endonuclease products, we analysed the ncRNA population. Among the ncRNAs accumulated in WT vs KO cells, we found mostly tRNA-derived fragments (tRFs) and second miRNAs. Differential sequence coverage identified tRFs from only few parental tRNAs, revealing a predominant cleavage at anticodon and D-loops at U/C (B1) and A (B2) sites. Selective tRNA cleavage was confirmed in vitro using the recombinant protein. Likewise, only few miRNAs were significantly more abundant in WT vs RNase2-KO cells. Complementarily, by screening of a tRF & tiRNA array, we identified an enriched population associated to RNase2 expression and RSV exposure. The results confirm the protein antiviral action and provide the first evidence of its cleavage selectivity on ncRNAs.Funding: Open Access Funding provided by Universitat Autònoma de Barcelona. This research was founded by Research work was supported by the Ministerio de Economía y Competitividad (SAF2015-66007P), co-financed by FEDER funds, by Agencia Estatal de Investigación (PID2019-106123GB-I00/AEI/10.13039/501100011033) and by Fundació La Marató de TV3 (2080310). LL and JL were supported by China Scholarship Council (CSC) predoctoral fellowshipsSpringer202220222022info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/53264http://dx.doi.org/10.1007/s00018-022-04229-xreponame:Repositorio Digital de la UPFinstname:Universitat Pompeu FabraInglésinfo:eu-repo/grantAgreement/ES/1PE/SAF2015-66007P© Lu Lu et al. 2022. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were madehttps://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repositori.upf.edu:10230/532642026-06-12T07:21:37Z
dc.title.none.fl_str_mv Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
title Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
spellingShingle Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
Lu, Lu
Genètica
Virus
Virus sincicial respiratori
Ribonucleases
title_short Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
title_full Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
title_fullStr Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
title_full_unstemmed Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
title_sort Selective cleavage of ncRNA and antiviral activity by RNase2/EDN in THP1-induced macrophages
dc.creator.none.fl_str_mv Lu, Lu
Li, Jiarui
Wei, Ranlei
Guidi, Irene
Cozzuto, Luca
Ponomarenko, Julia
Prats-Ejarque, Guillem
Boix, Ester
author Lu, Lu
author_facet Lu, Lu
Li, Jiarui
Wei, Ranlei
Guidi, Irene
Cozzuto, Luca
Ponomarenko, Julia
Prats-Ejarque, Guillem
Boix, Ester
author_role author
author2 Li, Jiarui
Wei, Ranlei
Guidi, Irene
Cozzuto, Luca
Ponomarenko, Julia
Prats-Ejarque, Guillem
Boix, Ester
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Genètica
Virus
Virus sincicial respiratori
Ribonucleases
topic Genètica
Virus
Virus sincicial respiratori
Ribonucleases
description RNase2 is the member of the RNaseA family most abundant in macrophages. Here, we knocked out RNase2 in THP-1 cells and analysed the response to Respiratory Syncytial Virus (RSV). RSV induced RNase2 expression, which significantly enhanced cell survival. Next, by cP-RNAseq sequencing, which amplifies the cyclic-phosphate endonuclease products, we analysed the ncRNA population. Among the ncRNAs accumulated in WT vs KO cells, we found mostly tRNA-derived fragments (tRFs) and second miRNAs. Differential sequence coverage identified tRFs from only few parental tRNAs, revealing a predominant cleavage at anticodon and D-loops at U/C (B1) and A (B2) sites. Selective tRNA cleavage was confirmed in vitro using the recombinant protein. Likewise, only few miRNAs were significantly more abundant in WT vs RNase2-KO cells. Complementarily, by screening of a tRF & tiRNA array, we identified an enriched population associated to RNase2 expression and RSV exposure. The results confirm the protein antiviral action and provide the first evidence of its cleavage selectivity on ncRNAs.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/53264
http://dx.doi.org/10.1007/s00018-022-04229-x
url http://hdl.handle.net/10230/53264
http://dx.doi.org/10.1007/s00018-022-04229-x
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/ES/1PE/SAF2015-66007P
dc.rights.none.fl_str_mv https://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:Repositorio Digital de la UPF
instname:Universitat Pompeu Fabra
instname_str Universitat Pompeu Fabra
reponame_str Repositorio Digital de la UPF
collection Repositorio Digital de la UPF
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