An analysis of the sequence requirements of EDEN-BP for specific RNA binding.
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.
Bonnet-Corven
Sylvie
S
UMR 6061 CNRS-Génétique et Développement, IFR 97, Université de Rennes 1, Faculté de Médecine, 2 Avenue Léon Bernard, CS 34317, 35043 Rennes Cedex, France.
Audic
Yann
Y
Omilli
Francis
F
Osborne
H Beverley
HB
D020413:3' Untranslated Regions / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs; D000818:Animals; D019281:Dimerization; D024202:Electrophoretic Mobility Shift Assay; D009865:Oocytes / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D013379:Substrate Specificity; D020798:Two-Hybrid System Techniques; D029867:Xenopus Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis / Q000196:embryology / Q000235:genetics / Q000378:metabolism
D020413:3' Untranslated Regions; C110462:EDEN-specific RNA-binding protein, Xenopus; D016601:RNA-Binding Proteins; D029867:Xenopus Proteins