The RNA binding domain of the Pumilio and FBF (PUF) proteins serves as a designable scaffold to recognize specific RNA elements, and the specificity to target RNA can be altered by introducing mutations with rational design. Thus, the RNA binding domains of PUF proteins are ideal scaffolds to regulate post-transcriptional processes of target mRNAs. Here, we use PUF scaffolds to regulate the activation and inhibition of mRNA translation in mammalian cells. We activate mRNA translation by tethering the fusion protein containing eukaryotic translation initiation factor 4E (eIF4E) and the PUF scaffold to the upstream of the target open reading frame (ORF), while repress mRNA translation by blocking the scanning of 43S preinitiation complex by localizing the PUF scaffold to the upstream of the target ORF. We have therefore demonstrated the strategy to regulate target mRNA translation in mammalian cells that could be potentially used to regulate the translation of specific endogenous mRNAs in vivo.

Regulation of mRNA translation using PUF protein scaffolds in mammalian cells

247th National Meeting (2014)

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