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PMID: 26578598 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Nucleotide modifications within bacterial messenger RNAs regulate their translation and are able to rewire the genetic code.

Nucleic acids research ·Vol. 44 ·No. 2 ·2016-01-29 ·Pages 852-62

Hoernes TP, Clementi N, Faserl K, Glasner H, Breuker K, Lindner H, Hüttenhofer A, Erlacher MD

Abstract

Nucleotide modifications within RNA transcripts are found in every organism in all three domains of life. 6-methyladeonsine (m(6)A), 5-methylcytosine (m(5)C) and pseudouridine (Ψ) are highly abundant nucleotide modifications in coding sequences of eukaryal mRNAs, while m(5)C and m(6)A modifications have also been discovered in archaeal and bacterial mRNAs. Employing in vitro translation assays, we systematically investigated the influence of nucleotide modifications on translation. We introduced m(5)C, m(6)A, Ψ or 2'-O-methylated nucleotides at each of the three positions within a codon of the bacterial ErmCL mRNA and analyzed their influence on translation. Depending on the respective nucleotide modification, as well as its position within a codon, protein synthesis remained either unaffected or was prematurely terminated at the modification site, resulting in reduced amounts of the full-length peptide. In the latter case, toeprint analysis of ribosomal complexes was consistent with stalling of translation at the modified codon. When multiple nucleotide modifications were introduced within one codon, an additive inhibitory effect on translation was observed. We also identified the m(5)C modification to alter the amino acid identity of the corresponding codon, when positioned at the second codon position. Our results suggest a novel mode of gene regulation by nucleotide modifications in bacterial mRNAs.

MeSH Terms
5-Methylcytosine/metabolism Adenosine/analogs & derivatives,genetics,metabolism Codon Escherichia coli/genetics Methyltransferases/genetics Protein Biosynthesis Pseudouridine/genetics,metabolism RNA/chemistry,metabolism RNA, Bacterial/genetics,metabolism RNA, Messenger/genetics,metabolism
Chemicals
Codon RNA, Bacterial RNA, Messenger Pseudouridine RNA 5-Methylcytosine N-methyladenosine Methyltransferases rRNA (adenosine-O-2'-)methyltransferase Adenosine
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Hoernes Thomas Philipp
Division of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, Austria.
Clementi Nina
Division of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, Austria.
Faserl Klaus
Division of Clinical Biochemistry, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, Austria.
Glasner Heidelinde
Institute of Organic Chemistry and Center for Molecular Biosciences (CMBI), University of Innsbruck, 6020 Innsbruck, Austria.
Breuker Kathrin
Institute of Organic Chemistry and Center for Molecular Biosciences (CMBI), University of Innsbruck, 6020 Innsbruck, Austria.
Lindner Herbert
Division of Clinical Biochemistry, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, Austria.
Hüttenhofer Alexander
Division of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, Austria alexander.huettenhofer@i-med.ac.at.
Erlacher Matthias David
Division of Genomics and RNomics, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, Austria matthias.erlacher@i-med.ac.at.
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkv1182
Email
alexander.huettenhofer@i-med.ac.at
matthias.erlacher@i-med.ac.at
Published
2016-01-29
Epub
2015-00-17
Pages
852-62
Language
English
Region
England
NLM ID
0411011
PMCID
PMC4737146
Subset
IM
Grants
Austrian Science Fund FWF · P 22658 · Austria
Austrian Science Fund FWF · P 27347 · Austria
External Links
PubMed source DOI: 10.1093/nar/gkv1182 PMC Full Text View journal details
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