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An extended dsRBD is required for post-transcriptional modification in human tRNAs

In tRNA, dihydrouridine is a conserved modified base generated by the post-transcriptional reduction of uridine. Formation of dihydrouridine 20, located in the D-loop, is catalyzed by dihydrouridine synthase 2 (Dus2). Human Dus2 (HsDus2) expression is upregulated in lung cancers, offering a growth a...

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Autores principales: Bou-Nader, Charles, Pecqueur, Ludovic, Bregeon, Damien, Kamah, Amina, Guérineau, Vincent, Golinelli-Pimpaneau, Béatrice, Guimarães, Beatriz G., Fontecave, Marc, Hamdane, Djemel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4627097/
https://www.ncbi.nlm.nih.gov/pubmed/26429968
http://dx.doi.org/10.1093/nar/gkv989
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author Bou-Nader, Charles
Pecqueur, Ludovic
Bregeon, Damien
Kamah, Amina
Guérineau, Vincent
Golinelli-Pimpaneau, Béatrice
Guimarães, Beatriz G.
Fontecave, Marc
Hamdane, Djemel
author_facet Bou-Nader, Charles
Pecqueur, Ludovic
Bregeon, Damien
Kamah, Amina
Guérineau, Vincent
Golinelli-Pimpaneau, Béatrice
Guimarães, Beatriz G.
Fontecave, Marc
Hamdane, Djemel
author_sort Bou-Nader, Charles
collection PubMed
description In tRNA, dihydrouridine is a conserved modified base generated by the post-transcriptional reduction of uridine. Formation of dihydrouridine 20, located in the D-loop, is catalyzed by dihydrouridine synthase 2 (Dus2). Human Dus2 (HsDus2) expression is upregulated in lung cancers, offering a growth advantage throughout its ability to interact with components of the translation apparatus and inhibit apoptosis. Here, we report the crystal structure of the individual domains of HsDus2 and their functional characterization. HsDus2 is organized into three major modules. The N-terminal catalytic domain contains the flavin cofactor involved in the reduction of uridine. The second module is the conserved α-helical domain known as the tRNA binding domain in HsDus2 homologues. It is connected via a flexible linker to an unusual extended version of a dsRNA binding domain (dsRBD). Enzymatic assays and yeast complementation showed that the catalytic domain binds selectively NADPH but cannot reduce uridine in the absence of the dsRBD. While in Dus enzymes from bacteria, plants and fungi, tRNA binding is essentially achieved by the α-helical domain, we showed that in HsDus2 this function is carried out by the dsRBD. This is the first reported case of a tRNA-modifying enzyme carrying a dsRBD used to bind tRNAs.
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spelling pubmed-46270972015-11-13 An extended dsRBD is required for post-transcriptional modification in human tRNAs Bou-Nader, Charles Pecqueur, Ludovic Bregeon, Damien Kamah, Amina Guérineau, Vincent Golinelli-Pimpaneau, Béatrice Guimarães, Beatriz G. Fontecave, Marc Hamdane, Djemel Nucleic Acids Res Nucleic Acid Enzymes In tRNA, dihydrouridine is a conserved modified base generated by the post-transcriptional reduction of uridine. Formation of dihydrouridine 20, located in the D-loop, is catalyzed by dihydrouridine synthase 2 (Dus2). Human Dus2 (HsDus2) expression is upregulated in lung cancers, offering a growth advantage throughout its ability to interact with components of the translation apparatus and inhibit apoptosis. Here, we report the crystal structure of the individual domains of HsDus2 and their functional characterization. HsDus2 is organized into three major modules. The N-terminal catalytic domain contains the flavin cofactor involved in the reduction of uridine. The second module is the conserved α-helical domain known as the tRNA binding domain in HsDus2 homologues. It is connected via a flexible linker to an unusual extended version of a dsRNA binding domain (dsRBD). Enzymatic assays and yeast complementation showed that the catalytic domain binds selectively NADPH but cannot reduce uridine in the absence of the dsRBD. While in Dus enzymes from bacteria, plants and fungi, tRNA binding is essentially achieved by the α-helical domain, we showed that in HsDus2 this function is carried out by the dsRBD. This is the first reported case of a tRNA-modifying enzyme carrying a dsRBD used to bind tRNAs. Oxford University Press 2015-10-30 2015-10-01 /pmc/articles/PMC4627097/ /pubmed/26429968 http://dx.doi.org/10.1093/nar/gkv989 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Nucleic Acid Enzymes
Bou-Nader, Charles
Pecqueur, Ludovic
Bregeon, Damien
Kamah, Amina
Guérineau, Vincent
Golinelli-Pimpaneau, Béatrice
Guimarães, Beatriz G.
Fontecave, Marc
Hamdane, Djemel
An extended dsRBD is required for post-transcriptional modification in human tRNAs
title An extended dsRBD is required for post-transcriptional modification in human tRNAs
title_full An extended dsRBD is required for post-transcriptional modification in human tRNAs
title_fullStr An extended dsRBD is required for post-transcriptional modification in human tRNAs
title_full_unstemmed An extended dsRBD is required for post-transcriptional modification in human tRNAs
title_short An extended dsRBD is required for post-transcriptional modification in human tRNAs
title_sort extended dsrbd is required for post-transcriptional modification in human trnas
topic Nucleic Acid Enzymes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4627097/
https://www.ncbi.nlm.nih.gov/pubmed/26429968
http://dx.doi.org/10.1093/nar/gkv989
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