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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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 (Ψ) 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′<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 (Ψ) 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′<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 |
work_keys_str_mv |
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