Detection and functional characterization of queuosine and m5C modification in RNA

tRNAs are subject to a variety of chemical modifications that modulate their function in translation. In this project, we have investigated two such modifications, queuosine (Q) and cytosine-5 methylation (m5C). Our earlier work in Schizosaccharomyces pombe showed that Dnmt2-dependent methylation of C38 in tRNA-Asp is strongly stimulated by Q, which replaces guanosine at the wobble position (Q34) in the anticodon of selected tRNAs. Interestingly, Q in eukaryotes originally comes from bacterial sources in nutrition and from the gut microflora, thus providing a connection between nutritional supply and translation in the host. In this proposal, we will develop novel methods to detect Q and m5C in RNA. This project will yield novel tools for the detection of these modifications and will further improve our understanding of the epitranscriptome in eukaryotes.