Fig. 4: Comparison between the 40S subunit in context of 80S and bound to Otu2.
From: Molecular basis for recognition and deubiquitination of 40S ribosomes by Otu2
a View on intersubunit bridge eB12 formed by the C-terminal tail (CTT) of 60S protein eL19 and the 40S ES6e as present in the non-rotated state 80S (PDB:6snt)12. b Same view as in a on the Otu2-bound 40S subunit. Instead of eL19, α4 of Otu2 binds to ES6e major groove; residues contained in basic patches (BP2) are indicated. Ub ubiquitin c, d views focusing on the Otu2 α1-α2 hairpin and α3 located close to intersubunit bridges eB13 (formed by 60S protein eL24 and 40S h44) and eB11 (formed by 60S H101/ES41 and eS8 of the 40S subunit) in 80S non-rotated state. The hairpin binds to h44 opposite of the eL24 binding site. When bound to the hairpin, h44 moves towards the left foot (h9ES3a) thereby compacting the rRNA binding pocket for the hairpin. e Western blot analysis (α-FLAG) of fractions after sucrose density gradient centrifugation. Lysates were analyzed from an otu2∆ubp3∆ yeast strain harboring a vector expressing wild-type (wt) Otu2 or indicated 3xFLAG-tagged Otu2 mutants. BP1-mut; K116A K117A R118A R119A, BP2-mut; R121A K123A R125A R129A. We obtained essentially the same results in at least three independent experiments. Original Blots are provided in the Source Data file.
