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Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle

Archaeal fibrillarin (aFib) is a well-characterized S-adenosyl methionine (SAM)-dependent RNA 2′-O-methyltransferase that is known to act in a large C/D ribonucleoprotein (RNP) complex together with Nop5 and L7Ae proteins and a box C/D guide RNA. In the reaction, the guide RNA serves to direct the m...

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Autores principales: Tomkuvienė, Miglė, Ličytė, Janina, Olendraitė, Ingrida, Liutkevičiūtė, Zita, Clouet-d'Orval, Béatrice, Klimašauskas, Saulius
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5558902/
https://www.ncbi.nlm.nih.gov/pubmed/28576826
http://dx.doi.org/10.1261/rna.059832.116
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author Tomkuvienė, Miglė
Ličytė, Janina
Olendraitė, Ingrida
Liutkevičiūtė, Zita
Clouet-d'Orval, Béatrice
Klimašauskas, Saulius
author_facet Tomkuvienė, Miglė
Ličytė, Janina
Olendraitė, Ingrida
Liutkevičiūtė, Zita
Clouet-d'Orval, Béatrice
Klimašauskas, Saulius
author_sort Tomkuvienė, Miglė
collection PubMed
description Archaeal fibrillarin (aFib) is a well-characterized S-adenosyl methionine (SAM)-dependent RNA 2′-O-methyltransferase that is known to act in a large C/D ribonucleoprotein (RNP) complex together with Nop5 and L7Ae proteins and a box C/D guide RNA. In the reaction, the guide RNA serves to direct the methylation reaction to a specific site in tRNA or rRNA by sequence complementarity. Here we show that a Pyrococcus abyssi aFib–Nop5 heterodimer can alone perform SAM-dependent 2′-O-methylation of 16S and 23S ribosomal RNAs in vitro independently of L7Ae and C/D guide RNAs. Using tritium-labeling, mass spectrometry, and reverse transcription analysis, we identified three in vitro 2′-O-methylated positions in the 16S rRNA of P. abyssi, positions lying outside of previously reported pyrococcal C/D RNP methylation sites. This newly discovered stand-alone activity of aFib–Nop5 may provide an example of an ancestral activity retained in enzymes that were recruited to larger complexes during evolution.
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spelling pubmed-55589022018-09-01 Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle Tomkuvienė, Miglė Ličytė, Janina Olendraitė, Ingrida Liutkevičiūtė, Zita Clouet-d'Orval, Béatrice Klimašauskas, Saulius RNA Report Archaeal fibrillarin (aFib) is a well-characterized S-adenosyl methionine (SAM)-dependent RNA 2′-O-methyltransferase that is known to act in a large C/D ribonucleoprotein (RNP) complex together with Nop5 and L7Ae proteins and a box C/D guide RNA. In the reaction, the guide RNA serves to direct the methylation reaction to a specific site in tRNA or rRNA by sequence complementarity. Here we show that a Pyrococcus abyssi aFib–Nop5 heterodimer can alone perform SAM-dependent 2′-O-methylation of 16S and 23S ribosomal RNAs in vitro independently of L7Ae and C/D guide RNAs. Using tritium-labeling, mass spectrometry, and reverse transcription analysis, we identified three in vitro 2′-O-methylated positions in the 16S rRNA of P. abyssi, positions lying outside of previously reported pyrococcal C/D RNP methylation sites. This newly discovered stand-alone activity of aFib–Nop5 may provide an example of an ancestral activity retained in enzymes that were recruited to larger complexes during evolution. Cold Spring Harbor Laboratory Press 2017-09 /pmc/articles/PMC5558902/ /pubmed/28576826 http://dx.doi.org/10.1261/rna.059832.116 Text en © 2017 Tomkuvienė et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Report
Tomkuvienė, Miglė
Ličytė, Janina
Olendraitė, Ingrida
Liutkevičiūtė, Zita
Clouet-d'Orval, Béatrice
Klimašauskas, Saulius
Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
title Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
title_full Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
title_fullStr Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
title_full_unstemmed Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
title_short Archaeal fibrillarin–Nop5 heterodimer 2′-O-methylates RNA independently of the C/D guide RNP particle
title_sort archaeal fibrillarin–nop5 heterodimer 2′-o-methylates rna independently of the c/d guide rnp particle
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5558902/
https://www.ncbi.nlm.nih.gov/pubmed/28576826
http://dx.doi.org/10.1261/rna.059832.116
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