Functional integration of a semi-synthetic azido-queuosine derivative into translation and a tRNA modification circuit
Substitution of the queuine nucleobase precursor preQ1 by an azide-containing derivative (azido-propyl-preQ1) led to incorporation of this clickable chemical entity into tRNA via transglycosylation in vitro as well as in vivo in Escherichia coli, Schizosaccharomyces pombe and human cells. The result...
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| Hauptverfasser: | , , , , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
28 September 2022
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| In: |
Nucleic acids research
Year: 2022, Jahrgang: 50, Heft: 18, Pages: 10785-10800 |
| ISSN: | 1362-4962 |
| DOI: | 10.1093/nar/gkac822 |
| Online-Zugang: | Verlag, kostenfrei, Volltext: https://doi.org/10.1093/nar/gkac822 Verlag, kostenfrei, Volltext: https://academic.oup.com/nar/article/50/18/10785/6725769?login=true 
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| Verfasserangaben: | Larissa Bessler, Navpreet Kaur, Lea-Marie Vogt, Laurin Flemmich, Carmen Siebenaller, Marie-Luise Winz, Francesca Tuorto, Ronald Micura, Ann E. Ehrenhofer-Murray and Mark Helm |
MARC
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| 100 | 1 | |a Bessler, Larissa Sabine |d 1995- |e VerfasserIn |0 (DE-588)1294135724 |0 (DE-627)1851105387 |4 aut | |
| 245 | 1 | 0 | |a Functional integration of a semi-synthetic azido-queuosine derivative into translation and a tRNA modification circuit |c Larissa Bessler, Navpreet Kaur, Lea-Marie Vogt, Laurin Flemmich, Carmen Siebenaller, Marie-Luise Winz, Francesca Tuorto, Ronald Micura, Ann E. Ehrenhofer-Murray and Mark Helm |
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| 520 | |a Substitution of the queuine nucleobase precursor preQ1 by an azide-containing derivative (azido-propyl-preQ1) led to incorporation of this clickable chemical entity into tRNA via transglycosylation in vitro as well as in vivo in Escherichia coli, Schizosaccharomyces pombe and human cells. The resulting semi-synthetic RNA modification, here termed Q-L1, was present in tRNAs on actively translating ribosomes, indicating functional integration into aminoacylation and recruitment to the ribosome. The azide moiety of Q-L1 facilitates analytics via click conjugation of a fluorescent dye, or of biotin for affinity purification. Combining the latter with RNAseq showed that TGT maintained its native tRNA substrate specificity in S. pombe cells. The semi-synthetic tRNA modification Q-L1 was also functional in tRNA maturation, in effectively replacing the natural queuosine in its stimulation of further modification of tRNAAsp with 5-methylcytosine at position 38 by the tRNA methyltransferase Dnmt2 in S. pombe. This is the first demonstrated in vivo integration of a synthetic moiety into an RNA modification circuit, where one RNA modification stimulates another. In summary, the scarcity of queuosinylation sites in cellular RNA, makes our synthetic q/Q system a 'minimally invasive' system for placement of a non-natural, clickable nucleobase within the total cellular RNA. | ||
| 650 | 4 | |a 5-Methylcytosine | |
| 650 | 4 | |a Azides | |
| 650 | 4 | |a Biotin | |
| 650 | 4 | |a Escherichia coli | |
| 650 | 4 | |a Fluorescent Dyes | |
| 650 | 4 | |a Humans | |
| 650 | 4 | |a Nucleoside Q | |
| 650 | 4 | |a RNA, Transfer | |
| 650 | 4 | |a RNA, Transfer, Asp | |
| 650 | 4 | |a Schizosaccharomyces | |
| 650 | 4 | |a tRNA Methyltransferases | |
| 700 | 1 | |a Kaur, Navpreet |e VerfasserIn |4 aut | |
| 700 | 1 | |a Vogt, Lea-Marie |e VerfasserIn |4 aut | |
| 700 | 1 | |a Flemmich, Laurin |e VerfasserIn |4 aut | |
| 700 | 1 | |a Siebenaller, Carmen |e VerfasserIn |4 aut | |
| 700 | 1 | |a Winz, Marie-Luise |e VerfasserIn |4 aut | |
| 700 | 1 | |a Tuorto, Francesca |e VerfasserIn |0 (DE-588)1122208154 |0 (DE-627)875231187 |0 (DE-576)481189092 |4 aut | |
| 700 | 1 | |a Micura, Ronald |e VerfasserIn |4 aut | |
| 700 | 1 | |a Ehrenhofer-Murray, Ann E. |e VerfasserIn |4 aut | |
| 700 | 1 | |a Helm, Mark |e VerfasserIn |4 aut | |
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