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

Nucleic Acids Res. 2002 Nov 1;30(21):4667-74. doi: 10.1093/nar/gkf586.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics
  • 3' Untranslated Regions / metabolism*
  • Amino Acid Motifs
  • Animals
  • Dimerization
  • Electrophoretic Mobility Shift Assay
  • Oocytes / metabolism
  • Protein Binding
  • Protein Structure, Quaternary
  • Protein Structure, Tertiary
  • RNA-Binding Proteins / chemistry*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Sequence Deletion
  • Substrate Specificity
  • Two-Hybrid System Techniques
  • Xenopus Proteins / chemistry*
  • Xenopus Proteins / genetics
  • Xenopus Proteins / metabolism*
  • Xenopus laevis* / embryology
  • Xenopus laevis* / genetics
  • Xenopus laevis* / metabolism

Substances

  • 3' Untranslated Regions
  • EDEN-specific RNA-binding protein, Xenopus
  • RNA-Binding Proteins
  • Xenopus Proteins