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Dinucleoside polyphosphates act as 5’-RNA caps in Escherichia coli

Oldřich Hudeček, Roberto Benoni, Martin Culka, Martin Hubálek, Lubomír Rulíšek, Josef Cvačka, View ORCID ProfileHana Cahová
doi: https://doi.org/10.1101/563817
Oldřich Hudeček
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nam. 2, 16610 Prague 6, Czech Republic
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Roberto Benoni
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nam. 2, 16610 Prague 6, Czech Republic
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Martin Culka
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nam. 2, 16610 Prague 6, Czech Republic
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Martin Hubálek
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nam. 2, 16610 Prague 6, Czech Republic
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Lubomír Rulíšek
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nam. 2, 16610 Prague 6, Czech Republic
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Josef Cvačka
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nam. 2, 16610 Prague 6, Czech Republic
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Hana Cahová
Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo nam. 2, 16610 Prague 6, Czech Republic
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  • ORCID record for Hana Cahová
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Abstract

Dinucleoside polyphosphates (NpnNs), discovered more than 50 years ago,1 are pleiotropic molecules present in almost all types of cells.2 It has been shown that their intracellular concentration can under stress conditions increase from the μM to mM range2,3. However, the cellular roles and mechanisms of action of NpnNs are still speculative4,5. They have never been considered as part of the RNA, even though they have similar chemical structures as already known RNA caps, such as the nicotinamide adenine dinucleotide (NAD)6–8 and 7-methylguanylate cap9. Here, we show that both methylated and non-methylated NpnNs serve as RNA caps in Escherichia coli (E. coli). NpnNs are excellent substrates for T7 and E. coli RNA polymerases (RNAP) and efficiently initiate transcription. Further, we demonstrate that the E. coli decapping enzyme RNA 5’ pyrophosphohydrolase (RppH) is able to remove the NpnNs-cap from the RNA. RppH was, however, not able to cleave the methylated forms of the NpnN-caps, suggesting that the methylation adds an additional layer to the RNA stability regulation. Our work introduces an original perspective on the chemical structure of RNA in prokaryotes and the function of RNA caps. This is the first evidence that small molecules like NpnNs can act in cells via their incorporation into RNA and influence the cellular metabolism.

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Posted March 01, 2019.
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Dinucleoside polyphosphates act as 5’-RNA caps in Escherichia coli
Oldřich Hudeček, Roberto Benoni, Martin Culka, Martin Hubálek, Lubomír Rulíšek, Josef Cvačka, Hana Cahová
bioRxiv 563817; doi: https://doi.org/10.1101/563817
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Dinucleoside polyphosphates act as 5’-RNA caps in Escherichia coli
Oldřich Hudeček, Roberto Benoni, Martin Culka, Martin Hubálek, Lubomír Rulíšek, Josef Cvačka, Hana Cahová
bioRxiv 563817; doi: https://doi.org/10.1101/563817

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