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Multiple links between 5-methylcytosine content of mRNA and translation

BACKGROUND: 5-Methylcytosine (m(5)C) is a prevalent base modification in tRNA and rRNA but it also occurs more broadly in the transcriptome, including in mRNA, where it serves incompletely understood molecular functions. In pursuit of potential links of m(5)C with mRNA translation, we performed poly...

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Autores principales: Schumann, Ulrike, Zhang, He-Na, Sibbritt, Tennille, Pan, Anyu, Horvath, Attila, Gross, Simon, Clark, Susan J., Yang, Li, Preiss, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7158060/
https://www.ncbi.nlm.nih.gov/pubmed/32293435
http://dx.doi.org/10.1186/s12915-020-00769-5
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author Schumann, Ulrike
Zhang, He-Na
Sibbritt, Tennille
Pan, Anyu
Horvath, Attila
Gross, Simon
Clark, Susan J.
Yang, Li
Preiss, Thomas
author_facet Schumann, Ulrike
Zhang, He-Na
Sibbritt, Tennille
Pan, Anyu
Horvath, Attila
Gross, Simon
Clark, Susan J.
Yang, Li
Preiss, Thomas
author_sort Schumann, Ulrike
collection PubMed
description BACKGROUND: 5-Methylcytosine (m(5)C) is a prevalent base modification in tRNA and rRNA but it also occurs more broadly in the transcriptome, including in mRNA, where it serves incompletely understood molecular functions. In pursuit of potential links of m(5)C with mRNA translation, we performed polysome profiling of human HeLa cell lysates and subjected RNA from resultant fractions to efficient bisulfite conversion followed by RNA sequencing (bsRNA-seq). Bioinformatic filters for rigorous site calling were devised to reduce technical noise. RESULTS: We obtained ~ 1000 candidate m(5)C sites in the wider transcriptome, most of which were found in mRNA. Multiple novel sites were validated by amplicon-specific bsRNA-seq in independent samples of either human HeLa, LNCaP and PrEC cells. Furthermore, RNAi-mediated depletion of either the NSUN2 or TRDMT1 m(5)C:RNA methyltransferases showed a clear dependence on NSUN2 for the majority of tested sites in both mRNAs and noncoding RNAs. Candidate m(5)C sites in mRNAs are enriched in 5′UTRs and near start codons and are embedded in a local context reminiscent of the NSUN2-dependent m(5)C sites found in the variable loop of tRNA. Analysing mRNA sites across the polysome profile revealed that modification levels, at bulk and for many individual sites, were inversely correlated with ribosome association. CONCLUSIONS: Our findings emphasise the major role of NSUN2 in placing the m(5)C mark transcriptome-wide. We further present evidence that substantiates a functional interdependence of cytosine methylation level with mRNA translation. Additionally, we identify several compelling candidate sites for future mechanistic analysis. SUPPLEMENTARY INFORMATION: Supplementary information accompanies this paper at 10.1186/s12915-020-00769-5.
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spelling pubmed-71580602020-04-21 Multiple links between 5-methylcytosine content of mRNA and translation Schumann, Ulrike Zhang, He-Na Sibbritt, Tennille Pan, Anyu Horvath, Attila Gross, Simon Clark, Susan J. Yang, Li Preiss, Thomas BMC Biol Research Article BACKGROUND: 5-Methylcytosine (m(5)C) is a prevalent base modification in tRNA and rRNA but it also occurs more broadly in the transcriptome, including in mRNA, where it serves incompletely understood molecular functions. In pursuit of potential links of m(5)C with mRNA translation, we performed polysome profiling of human HeLa cell lysates and subjected RNA from resultant fractions to efficient bisulfite conversion followed by RNA sequencing (bsRNA-seq). Bioinformatic filters for rigorous site calling were devised to reduce technical noise. RESULTS: We obtained ~ 1000 candidate m(5)C sites in the wider transcriptome, most of which were found in mRNA. Multiple novel sites were validated by amplicon-specific bsRNA-seq in independent samples of either human HeLa, LNCaP and PrEC cells. Furthermore, RNAi-mediated depletion of either the NSUN2 or TRDMT1 m(5)C:RNA methyltransferases showed a clear dependence on NSUN2 for the majority of tested sites in both mRNAs and noncoding RNAs. Candidate m(5)C sites in mRNAs are enriched in 5′UTRs and near start codons and are embedded in a local context reminiscent of the NSUN2-dependent m(5)C sites found in the variable loop of tRNA. Analysing mRNA sites across the polysome profile revealed that modification levels, at bulk and for many individual sites, were inversely correlated with ribosome association. CONCLUSIONS: Our findings emphasise the major role of NSUN2 in placing the m(5)C mark transcriptome-wide. We further present evidence that substantiates a functional interdependence of cytosine methylation level with mRNA translation. Additionally, we identify several compelling candidate sites for future mechanistic analysis. SUPPLEMENTARY INFORMATION: Supplementary information accompanies this paper at 10.1186/s12915-020-00769-5. BioMed Central 2020-04-15 /pmc/articles/PMC7158060/ /pubmed/32293435 http://dx.doi.org/10.1186/s12915-020-00769-5 Text en © The Author(s) 2020 Open AccessThis 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 made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Schumann, Ulrike
Zhang, He-Na
Sibbritt, Tennille
Pan, Anyu
Horvath, Attila
Gross, Simon
Clark, Susan J.
Yang, Li
Preiss, Thomas
Multiple links between 5-methylcytosine content of mRNA and translation
title Multiple links between 5-methylcytosine content of mRNA and translation
title_full Multiple links between 5-methylcytosine content of mRNA and translation
title_fullStr Multiple links between 5-methylcytosine content of mRNA and translation
title_full_unstemmed Multiple links between 5-methylcytosine content of mRNA and translation
title_short Multiple links between 5-methylcytosine content of mRNA and translation
title_sort multiple links between 5-methylcytosine content of mrna and translation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7158060/
https://www.ncbi.nlm.nih.gov/pubmed/32293435
http://dx.doi.org/10.1186/s12915-020-00769-5
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