CUG-BP1/CELF1 requires UGU-rich sequences for high-affinity binding
| Autor: | Julien Marquis, Olivier Danos, Luc Paillard, Christine Le Bec, H. Beverley Osborne, Bertrand Cosson, Yann Audic |
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| Přispěvatelé: | Centre de recherche et d'applications sur les thérapies géniques (CRATG), Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS)-Généthon, Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Association Francaise contre les Myopathies Association de la Recherche contre le Cancer (N°4791) Ministere de la Jeunesse, de l'Education Nationale et de la Recherche- ACI BCMS314 European Union DG XII biotechnology program (QLK3-CT-2000-00721) CNRS Genethon Rennes Metropole., Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Osborne, Howard Beverley, Gene Expression and Development, Université de Rennes (UR) |
| Jazyk: | angličtina |
| Rok vydání: | 2006 |
| Předmět: |
MESH: SELEX Aptamer Technique
Xenopus MESH: Aptamers Nucleotide RNA-binding protein Biosensing Techniques Xenopus Proteins Biochemistry CELF1 Protein 0302 clinical medicine Trinucleotide Repeats MESH: Reverse Transcriptase Polymerase Chain Reaction MESH: Animals MESH: Xenopus 3' Untranslated Regions MESH: Xenopus Proteins [SDV.BDD]Life Sciences [q-bio]/Development Biology 0303 health sciences MESH: Kinetics Reverse Transcriptase Polymerase Chain Reaction SELEX Aptamer Technique Life Sciences RNA-Binding Proteins MESH: 3' Untranslated Regions Aptamers Nucleotide Ligand (biochemistry) MESH: Surface Plasmon Resonance RNA splicing Female MESH: Biosensing Techniques Research Article congenital hereditary and neonatal diseases and abnormalities MESH: Trinucleotide Repeats Aptamer Biology Sensitivity and Specificity 03 medical and health sciences [SDV.BDD] Life Sciences [q-bio]/Development Biology Animals Humans RNA Messenger Binding site Molecular Biology 030304 developmental biology MESH: RNA Messenger Binding Sites MESH: Humans Cell Biology Surface Plasmon Resonance Molecular biology MESH: Sensitivity and Specificity Kinetics MESH: RNA-Binding Proteins MESH: Binding Sites MESH: Female 030217 neurology & neurosurgery Systematic evolution of ligands by exponential enrichment |
| Zdroj: | Biochemical Journal Biochemical Journal, 2006, 400 (2), pp.291-301. ⟨10.1042/BJ20060490⟩ Biochemical Journal, Portland Press, 2006, 400 (2), pp.291-301. ⟨10.1042/BJ20060490⟩ |
| ISSN: | 0264-6021 1470-8728 |
| DOI: | 10.1042/BJ20060490⟩ |
| Popis: | International audience; CUG-BP1 [CUG-binding protein 1 also called CELF (CUG-BP1 and ETR3 like factors) 1] is a human RNA-binding protein that has been implicated in the control of splicing and mRNA translation. The Xenopus homologue [EDEN-BP (embryo deadenylation element-binding protein)] is required for rapid deadenylation of certain maternal mRNAs just after fertilization. A variety of sequence elements have been described as target sites for these two proteins but their binding specificity is still controversial. Using a SELEX (systematic evolution of ligand by exponential enrichment) procedure and recombinant CUG-BP1 we selected two families of aptamers. Surface plasmon resonance and electrophoretic mobility-shift assays showed that these two families differed in their ability to bind CUG-BP1. Furthermore, the selected high-affinity aptamers form two complexes with CUG-BP1 in electrophoretic mobility assays whereas those that bind with low affinity only form one complex. The validity of the distinction between the two families of aptamers was confirmed by a functional in vivo deadenylation assay. Only those aptamers that bound CUG-BP1 with high affinity conferred deadenylation on a reporter mRNA. These high-affinity RNAs are characterized by a richness in UGU motifs. Using these binding site characteristics we identified the Xenopus maternal mRNA encoding the MAPK (mitogen-activated protein kinase) phosphatase (XCl100alpha) as a substrate for EDEN-BP. In conclusion, high-affinity CUG-BP1 binding sites are sequence elements at least 30 nucleotides in length that are enriched in combinations of U and G nucleotides and contain at least 4 UGU trinucleotide motifs. Such sequence elements are functionally competent to target an RNA for deadenylation in vivo. |
| Databáze: | OpenAIRE |
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