CUG-BP1/CELF1 requires UGU-rich sequences for high-affinity binding.

Détails

ID Serval
serval:BIB_1B4420D53754
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
CUG-BP1/CELF1 requires UGU-rich sequences for high-affinity binding.
Périodique
The Biochemical journal
Auteur(s)
Marquis Julien, Paillard Luc, Audic Yann, Cosson Bertrand, Danos Olivier, Le Bec Christine, Osborne H. Beverley
Statut éditorial
Publié
Date de publication
12/2006
Volume
400
Numéro
2
Pages
291-301
Langue
anglais
Résumé
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.
Mots-clé
Animals, Humans, Kinetics, Female, 3’ Untranslated Regions, Binding Sites, RNA, Messenger/genetics/metabolism, Reverse Transcriptase Polymerase Chain Reaction, Aptamers, Nucleotide/*genetics/*metabolism, Biosensing Techniques, CELF1 Protein, RNA-Binding Proteins/*genetics/*metabolism, SELEX Aptamer Technique, Sensitivity and Specificity, Surface Plasmon Resonance, Trinucleotide Repeats, Xenopus, Xenopus Proteins/*genetics/*metabolism
Pubmed
Création de la notice
19/02/2020 12:23
Dernière modification de la notice
19/06/2020 5:26
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