An analysis of the sequence requirements of EDEN-BP for specific RNA binding

• Published in: Nucleic acids research Vol. 30; no. 21; pp. 4667 - 74
• Main Authors: Bonnet-Corven, Sylvie, Audic, Yann, Omilli, Francis, Osborne, Howard Beverley
• Format: Journal Article
• Language: English
• Published: Oxford University Press 01.11.2002
• Subjects: 3' Untranslated Regions; Amino Acid Motifs; Animals; Development Biology; Dimerization; Electrophoretic Mobility Shift Assay; Life Sciences; Oocytes; Protein Binding; Protein Structure, Quaternary; Protein Structure, Tertiary; RNA-Binding Proteins; Sequence Deletion; Substrate Specificity; Two-Hybrid System Techniques; Xenopus laevis; Xenopus Proteins
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkf586

EDEN-BP (embryo deadenylation element-binding protein) binds specifically to the EDEN motif in the 3'-untranslated regions of maternal mRNAs and targets these mRNAs for deadenylation and translational repression in Xenopus laevis embryos. EDEN-BP contains three RNA recognition motifs (RRMs) and is related to the elav family of RNA-binding proteins. In the present study we show that the two N-terminal RRMs of EDEN-BP are necessary for the interaction with EDEN as well as a part of the linker region (between RRM2 and RRM3). Using a band shift assay we show that two different complexes are formed according to the size and, therefore, the functional nature of the EDEN motif. Finally, we show that EDEN-BP can form a dimer in a two-hybrid assay. Accordingly, we suggest that the functional configuration of EDEN-BP is a dimer.