High-Throughput Sequencing Reveals Circular Substrates for an Archaeal RNA ligase.

Hubert F Becker 1, 2 Alice Héliou 2, 3, 4 Kamel Djaout 2 Roxane Lestini 2 Mireille Regnier 3, 4 Hannu Myllykallio 2
3 AMIB - Algorithms and Models for Integrative Biology
LIX - Laboratoire d'informatique de l'École polytechnique [Palaiseau], LRI - Laboratoire de Recherche en Informatique, UP11 - Université Paris-Sud - Paris 11, Inria Saclay - Ile de France
Abstract : It is only recently that the abundant presence of circular RNAs (circRNAs) in all kingdoms of Life, including the hyperthermophilic archaeon Pyrococcus abyssi, has emerged. This led us to investigate the physiological significance of a previously observed weak intramolecular ligation activity of Pab1020 RNA ligase. Here we demonstrate that this enzyme, despite sharing significant sequence similarity with DNA ligases, is indeed an RNA-specific polynucleotide ligase efficiently acting on physiologically significant substrates. Using a combination of RNA immunoprecipitation assays and RNA-seq, our genome-wide studies revealed 133 individual circRNA loci in P. abyssi. The large majority of these loci interacted with Pab1020 in cells and circularization of selected C/D Box and 5S rRNA transcripts was confirmed biochemically. Altogether these studies revealed that Pab1020 is required for RNA circularization. Our results further suggest the functional speciation of an ancestral NTase domain and/or DNA ligase towards RNA ligase activity and prompt for further characterization of the widespread functions of circular RNAs in prokaryotes. Detailed insight into the cellular substrates of Pab1020 may facilitate the development of new biotechnological applications e.g. in ligation of preadenylated adaptors to RNA molecules.
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Journal articles
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https://hal.archives-ouvertes.fr/hal-01491132
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Submitted on : Thursday, March 16, 2017 - 1:36:00 PM
Last modification on : Wednesday, March 27, 2019 - 4:41:29 PM

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  • HAL Id : hal-01491132, version 1
  • PUBMED : 28277897

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Hubert F Becker, Alice Héliou, Kamel Djaout, Roxane Lestini, Mireille Regnier, et al.. High-Throughput Sequencing Reveals Circular Substrates for an Archaeal RNA ligase.. RNA Biology, Taylor & Francis, 2017. ⟨hal-01491132⟩

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