Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAs

RNA modifications are crucial factors for efficient protein synthesis. All classes of RNAs that are involved in translation are modified to different extents. Recently, mRNA modifications and their impact on gene regulation became a focus of interest because they can exert a variety of effects on th...

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Main Authors: Thomas Philipp Hoernes, David Heimdörfer, Daniel Köstner, Klaus Faserl, Felix Nußbaumer, Raphael Plangger, Christoph Kreutz, Herbert Lindner, Matthias David Erlacher
Format: Article
Language:English
Published: MDPI AG 2019-01-01
Series:Genes
Subjects:
Online Access:https://www.mdpi.com/2073-4425/10/2/84
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spelling doaj-24a4eefdbe6246ee9401bcc673fdde822020-11-25T00:27:21ZengMDPI AGGenes2073-44252019-01-011028410.3390/genes10020084genes10020084Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAsThomas Philipp Hoernes0David Heimdörfer1Daniel Köstner2Klaus Faserl3Felix Nußbaumer4Raphael Plangger5Christoph Kreutz6Herbert Lindner7Matthias David Erlacher8Division of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, AustriaDivision of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, AustriaDivision of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, AustriaDivision of Clinical Biochemistry, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Organic Chemistry and Center for Molecular Biosciences (CMBI), University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Organic Chemistry and Center for Molecular Biosciences (CMBI), University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Organic Chemistry and Center for Molecular Biosciences (CMBI), University of Innsbruck, 6020 Innsbruck, AustriaDivision of Clinical Biochemistry, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, AustriaDivision of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, AustriaRNA modifications are crucial factors for efficient protein synthesis. All classes of RNAs that are involved in translation are modified to different extents. Recently, mRNA modifications and their impact on gene regulation became a focus of interest because they can exert a variety of effects on the fate of mRNAs. mRNA modifications within coding sequences can either directly or indirectly interfere with protein synthesis. In order to investigate the roles of various natural occurring modified nucleotides, we site-specifically introduced them into the coding sequence of reporter mRNAs and subsequently translated them in HEK293T cells. The analysis of the respective protein products revealed a strong position-dependent impact of RNA modifications on translation efficiency and accuracy. Whereas a single 5-methylcytosine (m<sup>5</sup>C) or pseudouridine (&#936;) did not reduce product yields, <i>N</i><sup>1</sup>-methyladenosine (m<sup>1</sup>A) generally impeded the translation of the respective modified mRNA. An inhibitory effect of 2&#8242;<i>O</i>-methlyated nucleotides (Nm) and <i>N</i><sup>6</sup>-methyladenosine (m<sup>6</sup>A) was strongly dependent on their position within the codon. Finally, we could not attribute any miscoding potential to the set of mRNA modifications tested in HEK293T cells.https://www.mdpi.com/2073-4425/10/2/84mRNA modificationribosomedecodingtranslation
collection DOAJ
language English
format Article
sources DOAJ
author Thomas Philipp Hoernes
David Heimdörfer
Daniel Köstner
Klaus Faserl
Felix Nußbaumer
Raphael Plangger
Christoph Kreutz
Herbert Lindner
Matthias David Erlacher
spellingShingle Thomas Philipp Hoernes
David Heimdörfer
Daniel Köstner
Klaus Faserl
Felix Nußbaumer
Raphael Plangger
Christoph Kreutz
Herbert Lindner
Matthias David Erlacher
Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAs
Genes
mRNA modification
ribosome
decoding
translation
author_facet Thomas Philipp Hoernes
David Heimdörfer
Daniel Köstner
Klaus Faserl
Felix Nußbaumer
Raphael Plangger
Christoph Kreutz
Herbert Lindner
Matthias David Erlacher
author_sort Thomas Philipp Hoernes
title Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAs
title_short Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAs
title_full Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAs
title_fullStr Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAs
title_full_unstemmed Eukaryotic Translation Elongation is Modulated by Single Natural Nucleotide Derivatives in the Coding Sequences of mRNAs
title_sort eukaryotic translation elongation is modulated by single natural nucleotide derivatives in the coding sequences of mrnas
publisher MDPI AG
series Genes
issn 2073-4425
publishDate 2019-01-01
description RNA modifications are crucial factors for efficient protein synthesis. All classes of RNAs that are involved in translation are modified to different extents. Recently, mRNA modifications and their impact on gene regulation became a focus of interest because they can exert a variety of effects on the fate of mRNAs. mRNA modifications within coding sequences can either directly or indirectly interfere with protein synthesis. In order to investigate the roles of various natural occurring modified nucleotides, we site-specifically introduced them into the coding sequence of reporter mRNAs and subsequently translated them in HEK293T cells. The analysis of the respective protein products revealed a strong position-dependent impact of RNA modifications on translation efficiency and accuracy. Whereas a single 5-methylcytosine (m<sup>5</sup>C) or pseudouridine (&#936;) did not reduce product yields, <i>N</i><sup>1</sup>-methyladenosine (m<sup>1</sup>A) generally impeded the translation of the respective modified mRNA. An inhibitory effect of 2&#8242;<i>O</i>-methlyated nucleotides (Nm) and <i>N</i><sup>6</sup>-methyladenosine (m<sup>6</sup>A) was strongly dependent on their position within the codon. Finally, we could not attribute any miscoding potential to the set of mRNA modifications tested in HEK293T cells.
topic mRNA modification
ribosome
decoding
translation
url https://www.mdpi.com/2073-4425/10/2/84
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