Methylated mRNA Nucleotides as Regulators for Ribosomal Translation
Methylated RNA nucleotides were recently discovered to be highly abundant in RNAs. The effects of these methylations were mainly attributed to altered mRNA stabilities, protein-binding affinities, or RNA structures. The direct impact of RNA modifications on the performance of the ribosome has not been investigated so far. In this chapter, we describe an approach that allows introducing RNA modifications site-specifically into coding sequences of mRNAs and determining their effect on the translation machinery in a well-defined bacterial in vitro system.
Key wordsSplinted ligation In vitro translation RNA purification Peptide purification
This work was supported by the FWF (P 22658-B12 and P 28494-BBL to M.E.)
- 9.Hoernes TP, Erlacher MD (2016) Translating the epitranscriptome. WIREs RNA 2016. doi: 10.1002/wrna.1375
- 33.Bommer U, Burkhardt N, Jünemann R, Spahn CM, Triana-Alonso FJ, Nierhaus KH (1997) Ribosomes and polysomes. Subcellular fractionation—a practical approach. IRL Press, Washington, DC, pp 271–301Google Scholar