Ribosome profiling of Murine Embryonic Fibroblasts in the context of different Fbxo4 and hnRNPK status.

Background: SCF-Fbxo4 is suspected to regulate activity of RNA binding protein hnRNPK by non-degradive ubiquitylation. Research strategy: To determine the genomic output of SCF-Fbxo4 - hnRNPK interplay, murine embryonic fibroblasts (MEFs) with different status of hnRNPK/Fbxo4 were subjected to Ribosome profiling. Sequencing was performed on total RNA and RNA protected by ribosomes for each condition. Applying RNAi strategy hnRNPK was depleted in either wild type MEFs or MEFs Fbxo(-/-). MEFs conditioned in 4 different ways: wt- control (I), Fbxo4 knock-out (II), Fbxo4 knock-out + hnRNPK RNAi knock-down (III) and RNAi hnRNPK knock-down only (IV) were subjected to sequencing of total RNA and Ribosome protected fragment.