FICC-Seq: a method for enzyme-specified profiling of methyl-5-uridine in cellular RNA
- Publication type:
- Journal article
- Metadata:
-
- Autoren
- Jean-Michel Carter
- Warren Emmett
- Igor RDL Mozos
- Annika Kotter
- Mark Helm
- Jernej Ule
- Shobbir Hussain
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000496767900001&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1093/nar/gkz658
- eISSN
- 1362-4962
- Externe Identifier
- Clarivate Analytics Document Solution ID: JN2ZB
- PubMed Identifier: 31361898
- ISSN
- 0305-1048
- Ausgabe der Veröffentlichung
- 19
- Zeitschrift
- NUCLEIC ACIDS RESEARCH
- Paginierung
- E113 - +
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Titel
- FICC-Seq: a method for enzyme-specified profiling of methyl-5-uridine in cellular RNA
- Sub types
- Article
- Ausgabe der Zeitschrift
- 47
Data source: Web of Science (Lite)
- Other metadata sources:
-
- Abstract
- <jats:title>Abstract</jats:title><jats:p>Methyl-5-uridine (m5U) is one the most abundant non-canonical bases present in cellular RNA, and in yeast is found at position U54 of tRNAs where modification is catalysed by the methyltransferase Trm2. Although the mammalian enzymes that catalyse m5U formation are yet to be identified via experimental evidence, based on sequence homology to Trm2, two candidates currently exist, TRMT2A and TRMT2B. Here we developed a genome-wide single-nucleotide resolution mapping method, Fluorouracil-Induced-Catalytic-Crosslinking-Sequencing (FICC-Seq), in order to identify the relevant enzymatic targets. We demonstrate that TRMT2A is responsible for the majority of m5U present in human RNA, and that it commonly targets U54 of cytosolic tRNAs. By comparison to current methods, we show that FICC-Seq is a particularly robust method for accurate and reliable detection of relevant enzymatic target sites. Our associated finding of extensive irreversible TRMT2A-tRNA crosslinking in vivo following 5-Fluorouracil exposure is also intriguing, as it suggests a tangible mechanism for a previously suspected RNA-dependent route of Fluorouracil-mediated cytotoxicity.</jats:p>
- Autoren
- Jean-Michel Carter
- Warren Emmett
- Igor Rdl Mozos
- Annika Kotter
- Mark Helm
- Jernej Ule
- Shobbir Hussain
- DOI
- 10.1093/nar/gkz658
- eISSN
- 1362-4962
- ISSN
- 0305-1048
- Ausgabe der Veröffentlichung
- 19
- Zeitschrift
- Nucleic Acids Research
- Sprache
- en
- Online publication date
- 2019
- Paginierung
- e113 - e113
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Herausgeber
- Oxford University Press (OUP)
- Herausgeber URL
- http://dx.doi.org/10.1093/nar/gkz658
- Datum der Datenerfassung
- 2021
- Titel
- FICC-Seq: a method for enzyme-specified profiling of methyl-5-uridine in cellular RNA
- Ausgabe der Zeitschrift
- 47
Data source: Crossref
- Abstract
- Methyl-5-uridine (m5U) is one the most abundant non-canonical bases present in cellular RNA, and in yeast is found at position U54 of tRNAs where modification is catalysed by the methyltransferase Trm2. Although the mammalian enzymes that catalyse m5U formation are yet to be identified via experimental evidence, based on sequence homology to Trm2, two candidates currently exist, TRMT2A and TRMT2B. Here we developed a genome-wide single-nucleotide resolution mapping method, Fluorouracil-Induced-Catalytic-Crosslinking-Sequencing (FICC-Seq), in order to identify the relevant enzymatic targets. We demonstrate that TRMT2A is responsible for the majority of m5U present in human RNA, and that it commonly targets U54 of cytosolic tRNAs. By comparison to current methods, we show that FICC-Seq is a particularly robust method for accurate and reliable detection of relevant enzymatic target sites. Our associated finding of extensive irreversible TRMT2A-tRNA crosslinking in vivo following 5-Fluorouracil exposure is also intriguing, as it suggests a tangible mechanism for a previously suspected RNA-dependent route of Fluorouracil-mediated cytotoxicity.
- Addresses
- Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, BA2 7AY, UK.
- Autoren
- Jean-Michel Carter
- Warren Emmett
- Igor Rdl Mozos
- Annika Kotter
- Mark Helm
- Jernej Ule
- Shobbir Hussain
- DOI
- 10.1093/nar/gkz658
- eISSN
- 1362-4962
- Externe Identifier
- PubMed Identifier: 31361898
- PubMed Central ID: PMC6821191
- Funding acknowledgements
- Biotechnology and Biological Sciences Research Council: BB/N000749/1
- Biotechnology and Biosciences Research Council UK: BB/N000749/1
- Open access
- true
- ISSN
- 0305-1048
- Ausgabe der Veröffentlichung
- 19
- Zeitschrift
- Nucleic acids research
- Schlüsselwörter
- Humans
- Saccharomyces cerevisiae
- Yeasts
- Fluorouracil
- Deoxyribonucleases
- tRNA Methyltransferases
- Saccharomyces cerevisiae Proteins
- RNA
- RNA, Transfer
- Uridine
- Cell Survival
- HEK293 Cells
- High-Throughput Nucleotide Sequencing
- Sprache
- eng
- Medium
- Open access status
- Open Access
- Paginierung
- e113
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Publisher licence
- CC BY
- Datum der Datenerfassung
- 2019
- Titel
- FICC-Seq: a method for enzyme-specified profiling of methyl-5-uridine in cellular RNA.
- Sub types
- Research Support, Non-U.S. Gov't
- research-article
- Journal Article
- Ausgabe der Zeitschrift
- 47
Files
https://academic.oup.com/nar/article-pdf/47/19/e113/30314346/gkz658.pdf https://europepmc.org/articles/PMC6821191?pdf=render
Data source: Europe PubMed Central
- Abstract
- Methyl-5-uridine (m5U) is one the most abundant non-canonical bases present in cellular RNA, and in yeast is found at position U54 of tRNAs where modification is catalysed by the methyltransferase Trm2. Although the mammalian enzymes that catalyse m5U formation are yet to be identified via experimental evidence, based on sequence homology to Trm2, two candidates currently exist, TRMT2A and TRMT2B. Here we developed a genome-wide single-nucleotide resolution mapping method, Fluorouracil-Induced-Catalytic-Crosslinking-Sequencing (FICC-Seq), in order to identify the relevant enzymatic targets. We demonstrate that TRMT2A is responsible for the majority of m5U present in human RNA, and that it commonly targets U54 of cytosolic tRNAs. By comparison to current methods, we show that FICC-Seq is a particularly robust method for accurate and reliable detection of relevant enzymatic target sites. Our associated finding of extensive irreversible TRMT2A-tRNA crosslinking in vivo following 5-Fluorouracil exposure is also intriguing, as it suggests a tangible mechanism for a previously suspected RNA-dependent route of Fluorouracil-mediated cytotoxicity.
- Date of acceptance
- 2019
- Autoren
- Jean-Michel Carter
- Warren Emmett
- Igor Rdl Mozos
- Annika Kotter
- Mark Helm
- Jernej Ule
- Shobbir Hussain
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/31361898
- DOI
- 10.1093/nar/gkz658
- eISSN
- 1362-4962
- Externe Identifier
- PubMed Central ID: PMC6821191
- Funding acknowledgements
- Biotechnology and Biological Sciences Research Council: BB/N000749/1
- Ausgabe der Veröffentlichung
- 19
- Zeitschrift
- Nucleic Acids Res
- Schlüsselwörter
- Cell Survival
- Deoxyribonucleases
- Fluorouracil
- HEK293 Cells
- High-Throughput Nucleotide Sequencing
- Humans
- RNA
- RNA, Transfer
- Saccharomyces cerevisiae
- Saccharomyces cerevisiae Proteins
- Uridine
- Yeasts
- tRNA Methyltransferases
- Sprache
- eng
- Country
- England
- Paginierung
- e113
- PII
- 5541094
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2019
- Titel
- FICC-Seq: a method for enzyme-specified profiling of methyl-5-uridine in cellular RNA.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Ausgabe der Zeitschrift
- 47
Data source: PubMed
- Beziehungen:
- Property of