Molecular determinants of metazoan tricRNA biogenesis
| Autor: | Casey A. Schmidt, A. Gregory Matera, Alicia Bao, Anita K. Hopper, Joseph D. Giusto |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
RNA Splicing
TRNA processing Saccharomyces cerevisiae Biology 03 medical and health sciences 0302 clinical medicine RNA Transfer Endoribonucleases RNA Precursors RNA and RNA-protein complexes Genetics Animals Humans Nucleotide Motifs 030304 developmental biology chemistry.chemical_classification 0303 health sciences DNA ligase Intron RNA RNA Circular Introns Cell biology chemistry 030220 oncology & carcinogenesis RNA splicing Transfer RNA Endonuclease complex Drosophila Biogenesis |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkz311 |
| Popis: | Mature tRNAs are generated by multiple post-transcriptional processing steps, which can include intron removal. Recently, we discovered a new class of circular non-coding RNAs in metazoans, called tRNA intronic circular (tric)RNAs. To investigate the mechanism of tricRNA biogenesis, we generated constructs that replace native introns of human and fruit fly tRNA genes with the Broccoli fluorescent RNA aptamer. Using these reporters, we identified cis-acting elements required for tricRNA formation in vivo. Disrupting a conserved base pair in the anticodon-intron helix dramatically reduces tricRNA levels. Although the integrity of this base pair is necessary for proper splicing, it is not sufficient. In contrast, strengthening weak bases in the helix also interferes with splicing and tricRNA production. Furthermore, we identified trans-acting factors important for tricRNA biogenesis, including several known tRNA processing enzymes such as the RtcB ligase and components of the TSEN endonuclease complex. Depletion of these factors inhibits Drosophila tRNA intron circularization. Notably, RtcB is missing from fungal genomes and these organisms normally produce linear tRNA introns. Here, we show that in the presence of ectopic RtcB, yeast lacking the tRNA ligase Rlg1/Trl1 are converted into producing tricRNAs. In summary, our work characterizes the major players in eukaryotic tricRNA biogenesis. |
| Databáze: | OpenAIRE |
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