Proteomics

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Inosine induces context-dependent recoding and translational stalling

ABSTRACT: To test how inosine in a codon is translated, we synthesized short reporter-transcripts coding for an N-terminal Flag-tag and a test-peptide containing inosine in one defined codon. All codons containing inosines in at least one position were tested. Codons that would lead to ambiguous translation products were omitted. The transcripts were generated using in vitro transcription with ITP instead of GTP and subsequently in vitro translated using rabbit reticulocyte lysate and resulting peptides were analyzed by LC-MS/MS. The templates for in vitro transcription were generated using linearized DNA plasmids.

INSTRUMENT(S): Q Exactive HF-X, Q Exactive HF

ORGANISM(S): Synthetic Construct

SUBMITTER: Konstantin Licht  

LAB HEAD: Michael F. Jantsch

PROVIDER: PXD010329 | Pride | 2018-11-06

REPOSITORIES: Pride

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Inosine induces context-dependent recoding and translational stalling.

Licht Konstantin K   Hartl Markus M   Amman Fabian F   Anrather Dorothea D   Janisiw Michael P MP   Jantsch Michael F MF  

Nucleic acids research 20190101 1

RNA modifications are present in all classes of RNAs. They control the fate of mRNAs by affecting their processing, translation, or stability. Inosine is a particularly widespread modification in metazoan mRNA arising from deamination of adenosine catalyzed by the RNA-targeting adenosine deaminases ADAR1 or ADAR2. Inosine is commonly thought to be interpreted as guanosine by cellular machines and during translation. Here, we systematically test ribosomal decoding using mass spectrometry. We show  ...[more]

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