Fig. 3: NAC facilitates co-translational acetylation by NAA40 and coordination with METAP1.
From: NAA40 and NAC cooperate in co-translational histone acetylation in humans
a In vitro binding assay of NAA40 and NAA40S with NAC and 80S. SDS-PAGE gel of input and elution fractions stained with Coomassie. b Schematic comparison between NAA40 and NAA40S (top). Molecular model of the first 21 amino acids exclusive to full-length NAA40 with positively charged amino acids interacting with rRNA shown (bottom). c Cartoon representation showing the recruitment of NAA40 to the ribosome by NAC via the NACA-UBA domain. d Western blot analysis of histone extracts from HCT116 Scramble, NACA-KD and NAA40 knock-out (KO) cells, using antibodies against N-terminal acetylation (Nt-Ac), its antagonistic mark H4/H2AS1Ph and H4 as a loading control. e Western blot analysis of histone extracts from HCT116 scramble, NACA-KD transduced with FLAG-NACA (WT) or FLAG-NACA (ΔUBA) siRNA resistant plasmids, using antibody against NAA40 antagonistic mark H4/H2AS1ph and H2A as a loading control. f Quantification of western blot signals H4/H2AS1Ph relative to H2A from (e). All statistical analyses were performed using unpaired One-Way ANOVA (ns no significance, **P < 0.01) (bottom), n = 3 biological replicates/group. P values up to 5 digits are shown. Error bars represent standard error of mean (SEM). g Overlay of ribosome bound NAA40-NAC model with ribosome bound METAP1 (PDB: 8P2K), showing no steric clashes. The question mark indicates that based on the overlay the potential for co-assembly of METAP1 with NAA40-NAC on the ribosome is given and tested in (h). h In vitro binding assay of NAA40 and METAP1 with NAC and ribosomes. SDS-PAGE gels of input and elution fractions stained with Coomassie. Source data are provided as a Source data file.
