Binding specificity and mRNA targets of a C. elegans PUF protein, FBF-1

Abstract

Sequence-specific RNA–protein interactions underlie regulation of many mRNAs. Here we analyze the RNA sequence specificity of Caenorhabditis elegans FBF-1, a founding member of the PUF protein family. Like other PUF proteins, FBF-1 binds to the 3′ UTR of target mRNAs and decreases expression of those target genes. Here, we show that FBF-1 and its close relative, FBF-2, bind with similar affinity to multiple RNA sites. We use mutagenesis and in vivo selection experiments to identify nucleotides that are essential for FBF-1 binding. The binding elements comprise a “core” central region and flanking sequences. The core region is similar but distinct from the binding sites of other PUF proteins. We combine the identification of binding elements with informatics to predict new FBF-1 binding sites in a C. elegans 3′ UTR database. These data identify a set of new candidate mRNA targets of FBF-1 and FBF-2.

Keywords

• RNA/protein interactions
• PUF protein
• three-hybrid system
• FBF