Study of the translation initiation by Selective TCP-seq in the context of eIF1-eIF4G1 inhibitor

Start codon recognition by the 48S complex is a critical step in translation. However, understanding the in vivo initiation and its regulation at a global scale is limited. To better understand the mechanism in vivo, we have screened for small molecules that specifically inhibit the function of eIF1-eIF4G1 interaction. We performed Selective-48S footprinting against eIF1 (HA-tagged), eIF2a (HA-tagged), eIF4G1 and eIF3c to answer questions regarding the function of that interaction in the context of the scanning and initiating 48S ribosome. Overall design: Selective 48S footprinting (TCP-seq) of eIF4G1, HAeIF1,HA eIF2a, eIF3 from DMSO or i14G1-12 treated cells in HEK 293T cells.