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A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets

Mobile group II introns are site-specific retrotransposable elements abundant in bacterial and organellar genomes. They are composed of a large and highly structured ribozyme and an intron-encoded reverse transcriptase that binds tightly to its intron to yield a ribonucleoprotein (RNP) particle. Dur...

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Autores principales: Monachello, Dario, Lauraine, Marc, Gillot, Sandra, Michel, François, Costa, Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8643678/
https://www.ncbi.nlm.nih.gov/pubmed/34791436
http://dx.doi.org/10.1093/nar/gkab1031
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author Monachello, Dario
Lauraine, Marc
Gillot, Sandra
Michel, François
Costa, Maria
author_facet Monachello, Dario
Lauraine, Marc
Gillot, Sandra
Michel, François
Costa, Maria
author_sort Monachello, Dario
collection PubMed
description Mobile group II introns are site-specific retrotransposable elements abundant in bacterial and organellar genomes. They are composed of a large and highly structured ribozyme and an intron-encoded reverse transcriptase that binds tightly to its intron to yield a ribonucleoprotein (RNP) particle. During the first stage of the mobility pathway, the intron RNA catalyses its own insertion directly into the DNA target site. Recognition of the proper target rests primarily on multiple base-pairing interactions between the intron RNA and the target DNA, while the protein makes contacts with only a few target positions by yet-unidentified mechanisms. Using a combination of comparative sequence analyses and in vivo mobility assays we demonstrate the existence of a new base-pairing interaction named EBS2a–IBS2a between the intron RNA and its DNA target site. This pairing adopts a Watson–Crick geometry and is essential for intron mobility, most probably by driving unwinding of the DNA duplex. Importantly, formation of EBS2a–IBS2a also requires the reverse transcriptase enzyme which stabilizes the pairing in a non-sequence-specific manner. In addition to bringing to light a new structural device that allows subgroup IIB1 and IIB2 introns to invade their targets with high efficiency and specificity our work has important implications for the biotechnological applications of group II introns in bacterial gene targeting.
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spelling pubmed-86436782021-12-06 A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets Monachello, Dario Lauraine, Marc Gillot, Sandra Michel, François Costa, Maria Nucleic Acids Res Nucleic Acid Enzymes Mobile group II introns are site-specific retrotransposable elements abundant in bacterial and organellar genomes. They are composed of a large and highly structured ribozyme and an intron-encoded reverse transcriptase that binds tightly to its intron to yield a ribonucleoprotein (RNP) particle. During the first stage of the mobility pathway, the intron RNA catalyses its own insertion directly into the DNA target site. Recognition of the proper target rests primarily on multiple base-pairing interactions between the intron RNA and the target DNA, while the protein makes contacts with only a few target positions by yet-unidentified mechanisms. Using a combination of comparative sequence analyses and in vivo mobility assays we demonstrate the existence of a new base-pairing interaction named EBS2a–IBS2a between the intron RNA and its DNA target site. This pairing adopts a Watson–Crick geometry and is essential for intron mobility, most probably by driving unwinding of the DNA duplex. Importantly, formation of EBS2a–IBS2a also requires the reverse transcriptase enzyme which stabilizes the pairing in a non-sequence-specific manner. In addition to bringing to light a new structural device that allows subgroup IIB1 and IIB2 introns to invade their targets with high efficiency and specificity our work has important implications for the biotechnological applications of group II introns in bacterial gene targeting. Oxford University Press 2021-11-17 /pmc/articles/PMC8643678/ /pubmed/34791436 http://dx.doi.org/10.1093/nar/gkab1031 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Nucleic Acid Enzymes
Monachello, Dario
Lauraine, Marc
Gillot, Sandra
Michel, François
Costa, Maria
A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets
title A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets
title_full A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets
title_fullStr A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets
title_full_unstemmed A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets
title_short A new RNA–DNA interaction required for integration of group II intron retrotransposons into DNA targets
title_sort new rna–dna interaction required for integration of group ii intron retrotransposons into dna targets
topic Nucleic Acid Enzymes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8643678/
https://www.ncbi.nlm.nih.gov/pubmed/34791436
http://dx.doi.org/10.1093/nar/gkab1031
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