Bacillus subtilis RNA deprotection enzyme RppH recognizes guanosine in the second position of its substrates
| Autor: | Jérémie Piton, Françoise Debart, Jean-Jacques Vasseur, Carine Tisné, Inès Li de la Sierra-Gallay, Valéry Larue, Ciarán Condon, Olivier Pellegrini, Yann Thillier, Audrey Dorléans |
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| Přispěvatelé: | Unité Propre de Recherche 9073, Institut de Biologie Physico-Chimique, Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM), Université Paris Descartes - Paris 5 (UPD5) - Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM) - Université de Montpellier (UM) - Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences RNA Stability Multidisciplinary biology Oligonucleotide [CHIM.ORGA]Chemical Sciences/Organic chemistry 030302 biochemistry & molecular biology RNA Guanosine Bacillus subtilis Biological Sciences Guanosine triphosphate [CHIM.ORGA] Chemical Sciences/Organic chemistry biology.organism_classification Exoribonucleases 03 medical and health sciences chemistry.chemical_compound chemistry Biochemistry Nucleotide 030304 developmental biology |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2013, 110 (22), pp.8858-8863. ⟨10.1073/pnas.1221510110⟩ Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2013, 110 (22), pp.8858-8863 |
| ISSN: | 0027-8424 1091-6490 |
| DOI: | 10.1073/pnas.1221510110⟩ |
| Popis: | The initiation of mRNA degradation often requires deprotection of its 5′ end. In eukaryotes, the 5′-methylguanosine (cap) structure is principally removed by the Nudix family decapping enzyme Dcp2, yielding a 5′-monophosphorylated RNA that is a substrate for 5′ exoribonucleases. In bacteria, the 5′-triphosphate group of primary transcripts is also converted to a 5′ monophosphate by a Nudix protein called RNA pyrophosphohydrolase (RppH), allowing access to both endo- and 5′ exoribonucleases. Here we present the crystal structures of Bacillus subtilis RppH (BsRppH) bound to GTP and to a triphosphorylated dinucleotide RNA. In contrast to Bdellovibrio bacteriovorus RppH, which recognizes the first nucleotide of its RNA targets, the B. subtilis enzyme has a binding pocket that prefers guanosine residues in the second position of its substrates. The identification of sequence specificity for RppH in an internal position was a highly unexpected result. NMR chemical shift mapping in solution shows that at least three nucleotides are required for unambiguous binding of RNA. Biochemical assays of BsRppH on RNA substrates with single-base–mutation changes in the first four nucleotides confirm the importance of guanosine in position two for optimal enzyme activity. Our experiments highlight important structural and functional differences between BsRppH and the RNA deprotection enzymes of distantly related bacteria. |
| Databáze: | OpenAIRE |
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