Fig. 2: SBDS and EFL1 catalyze GTP-dependent release of rebound eIF6 from mature cytoplasmic 60S ribosomal subunits.
From: eIF6 rebinding dynamically couples ribosome maturation and translation
a Sucrose gradient sedimentation of c-Kit+ bone marrow cell extracts (without cycloheximide) lysed in 20 mM Hepes pH 7.5, 2 mM Mg(CH3COO)2, 500 mM KCl and incubated for 10 min at 37 °C to allow eIF6 rebinding. eIF6 was detected by immunoblotting. Data are representative of two independent experiments. b Schematic overview of in vitro eIF6 release assay. See “Methods” section for further details. c Sucrose gradient sedimentation of reconstituted eIF6 release reaction mixes. Immunoblotting was used to detect eIF6. The ratio of 80S monosomes to 60S subunits is indicated. Shown is a representative experiment out of a total of two independent experiments.
