Figure 7.

Sel-TCP-Seq Detects Co-translational Assembly within the MFC in Yeast

(A) Co-translational interactions between a tagged “bait” FOI (e.g., FLAG-tagged eIF3a) and its nascent assembly “target” (e.g., yeast eIF3b via its RNA recognition motif [RRM]) lead to a characteristic increase of selected 80S FP coverage within the assembly partner CDS.

(B–F) FP coverage tracks for selected (black; bait protein indicated on top of the left axis) and unselected 80S FP (gray; right axis) across mRNAs encoding target proteins within the yeast multifactor complex (MFC): eIF3b (B and D), eIF3g (C), eIF3a (E), and eIF2β (F). Domain structures of target MFC subunits are shown under the graphs. Dotted lines indicate the selected 80S FP coverage increase. Tracks from replicate 1 are shown, and average fold induction from 4 or 2 replicates (replicates 1–4 or 1+2) with standard deviation is given for eIF3a-FLAG or eIF3c-FLAG as bait, respectively. Fold induction of the 80S Sel-TCP-seq coverage was normalized to that seen with unselected 80S.

(G) Speculative MFC assembly model. Question marks indicate an unknown mode of factor assembly.