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Structural determination in AUF1 required for high affinity binding to A + U-rich elements
C T DeMaria1, Y Sun, B J Wagner
1Department of Microbiology and Immunology, Bowman Gray School of Medicine of Wake Forest University, Winston-Salem, NC 27157-1064, USA.
Nucleic Acids Symposium Series
|January 1, 1997
Summary
The RNA-binding protein AUF1 (AU-rich element binding protein 1) binds messenger RNAs containing AU-rich elements. Highest affinity binding requires specific protein domains and multimerization, impacting mRNA degradation.
Area of Science:
- Molecular Biology
- RNA Biology
- Protein-RNA Interactions
Background:
- AUF1 (AU-rich element binding protein 1) is a key RNA-binding protein.
- It interacts with AU-rich elements (AREs) found in the 3' untranslated regions of specific mRNAs.
- This interaction is critical for regulating the stability and degradation of mRNAs encoding cytokines, oncoproteins, and G protein-coupled receptors.
Purpose of the Study:
- To identify the specific domains of AUF1 responsible for high-affinity binding to AREs.
- To elucidate the role of protein structure and interactions in AUF1's ARE-binding activity.
Main Methods:
- Analysis of AUF1 mutants with altered domain structures.
- Assessment of binding affinities of wild-type and mutant AUF1 to AREs.
- Investigation of AUF1 oligomerization states during ARE binding.
Main Results:
- Both RNA recognition motifs (RRMs) of AUF1 are necessary but not sufficient for high-affinity ARE binding.
- Optimal binding requires an N-terminal alanine/glycine-rich region and a C-terminal glutamine-rich region.
- AUF1 dimerization, mediated by the N-terminus, is essential, and the protein binds AREs as a hexamer.
Conclusions:
- High-affinity binding of AUF1 to AREs is a complex process.
- It depends not only on RNA recognition motifs but also on specific N- and C-terminal regions.
- Protein-protein interactions and multimerization into a hexameric complex are crucial for AUF1's regulatory function.