meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2020 |
| Outros Autores: | , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10451/47290 |
Resumo: | © 2020 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. |
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meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation© 2020 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.The fates of RNA species in a cell are controlled by ribonucleases, which degrade them by exploiting the universal structural 2'-OH group. This phenomenon plays a key role in numerous transformative technologies, for example, RNA interference and CRISPR/Cas13-based RNA editing systems. These approaches, however, are genetic or oligomer-based and so have inherent limitations. This has led to interest in the development of small molecules capable of degrading nucleic acids in a targeted manner. Here we describe click-degraders, small molecules that can be covalently attached to RNA species through click-chemistry and can degrade them, that are akin to ribonucleases. By using these molecules, we have developed the meCLICK-Seq (methylation CLICK-degradation Sequencing) a method to identify RNA modification substrates with high resolution at intronic and intergenic regions. The method hijacks RNA methyltransferase activity to introduce an alkyne, instead of a methyl, moiety on RNA. Subsequent copper(I)-catalyzed azide-alkyne cycloaddition reaction with the click-degrader leads to RNA cleavage and degradation exploiting a mechanism used by endogenous ribonucleases. Focusing on N6-methyladenosine (m6A), meCLICK-Seq identifies methylated transcripts, determines RNA methylase specificity, and reliably maps modification sites in intronic and intergenic regions. Importantly, we show that METTL16 deposits m6A to intronic polyadenylation (IPA) sites, which suggests a potential role for METTL16 in IPA and, in turn, splicing. Unlike other methods, the readout of meCLICK-Seq is depletion, not enrichment, of modified RNA species, which allows a comprehensive and dynamic study of RNA modifications throughout the transcriptome, including regions of low abundance. The click-degraders are highly modular and so may be exploited to study any RNA modification and design new technologies that rely on RNA degradation.We thank UKRI (BBSRC DTP scholarships to S.M. and H.K.C) and the Jardine Foundation and Cambridge Trust (Ph.D. scholarship to M.E.H.). K.T. was funded by a Wellcome Trust Sir Henry Wellcome Fellowship (grant reference RG94424). G.S.V. is funded by a Cancer Research UK Senior Cancer Fellowship (C22324/A23015). Research in the Shi lab is supported by Ludwig Institute for Cancer Research. Y.S. is American Cancer Society Research Professor. G.J.L.B. is a Royal Society University Research Fellow (URF\R\180019) and an FCT Investigator (IF/00624/2015).ACS PublicationsRepositório da Universidade de LisboaMikutis, SigitasGu, MuxinSendinc, ErdemHazemi, Madoka EKiely-Collins, HannahAspris, DemetriosVassiliou, George SShi, YangTzelepis, KonstantinosBernardes, Gonçalo J. L.2021-04-08T15:02:25Z2020-12-232020-12-23T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/47290engACS Cent Sci. 2020 Dec 23;6(12):2196-220810.1021/acscentsci.0c010942374-7951info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T16:50:07Zoai:repositorio.ul.pt:10451/47290Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T21:59:22.326381Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
| dc.title.none.fl_str_mv |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation |
| title |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation |
| spellingShingle |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation Mikutis, Sigitas |
| title_short |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation |
| title_full |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation |
| title_fullStr |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation |
| title_full_unstemmed |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation |
| title_sort |
meCLICK-Seq, substrate-hijacking and RNA degradation strategy for the study of RNA methylation |
| author |
Mikutis, Sigitas |
| author_facet |
Mikutis, Sigitas Gu, Muxin Sendinc, Erdem Hazemi, Madoka E Kiely-Collins, Hannah Aspris, Demetrios Vassiliou, George S Shi, Yang Tzelepis, Konstantinos Bernardes, Gonçalo J. L. |
| author_role |
author |
| author2 |
Gu, Muxin Sendinc, Erdem Hazemi, Madoka E Kiely-Collins, Hannah Aspris, Demetrios Vassiliou, George S Shi, Yang Tzelepis, Konstantinos Bernardes, Gonçalo J. L. |
| author2_role |
author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Repositório da Universidade de Lisboa |
| dc.contributor.author.fl_str_mv |
Mikutis, Sigitas Gu, Muxin Sendinc, Erdem Hazemi, Madoka E Kiely-Collins, Hannah Aspris, Demetrios Vassiliou, George S Shi, Yang Tzelepis, Konstantinos Bernardes, Gonçalo J. L. |
| description |
© 2020 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. |
| publishDate |
2020 |
| dc.date.none.fl_str_mv |
2020-12-23 2020-12-23T00:00:00Z 2021-04-08T15:02:25Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10451/47290 |
| url |
http://hdl.handle.net/10451/47290 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
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ACS Cent Sci. 2020 Dec 23;6(12):2196-2208 10.1021/acscentsci.0c01094 2374-7951 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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ACS Publications |
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ACS Publications |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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