Reconstitution of Targeted Deadenylation by the Ccr4-Not Complex and the YTH Domain Protein Mmi1.

摘要: Ccr4-Not is a conserved protein complex that shortens the 3' poly(A) tails of eukaryotic mRNAs to regulate transcript stability and translation into proteins. RNA-binding proteins are thought to facilitate recruitment certain mRNAs, but lack an in-vitro-reconstituted system has slowed progress in understanding mechanistic details this specificity. Here, we generate fully recombinant removes from RNA substrates. The intact more active than exonucleases alone intrinsic preference for RNAs. Mmi1 highly abundant preparations native Ccr4-Not. We demonstrate high-affinity interaction between Mmi1. Using in vitro assays, show accelerates deadenylation target Together, our results support model whereby both RNA-binding proteins sequence context influence rate regulate gene expression.