Fig. 5: ALKBH8 regulates the translation elongation of KRAS.

a KEGG analysis of genes enriched with A-ending codons and exhibiting increased ribosome density following ALKBH8 knockout. 2 independent sequencing experiments, P values were calculated using DAVID (modified Fisher’s exact test with Benjamini–Hochberg correction). b Transcriptomic distribution of the percentage of 5A-ending codons. The percentage of A-ending codons for KRAS and Kras gene was highlighted. c Immunoblot analysis of KRAS expression in CTL and A8-KO HCT116 cells. d KRAS mRNA levels in control, A8-KO HCT116 cells. Mean ± SEM, 3 biological replicates, one-way ANOVA with Tukey’s multiple comparisons test. e Immunoblot analysis of 3 pairs of tumors from Apc ^min/+;Alkbh8^CTL and Apc ^min/+;Alkbh8 ^cKO mice. f Immunoblot analysis of 3 pairs of tumors from Alkbh8^CTL and Alkbh8 ^cKO mice of AOM/DSS induction. g Schematic of synonymous mutation of KRAS gene. h The expression of wild-type KRAS gene and its synonymous mutant in CTL and A8-KO cells. i The mRNA level of KRAS in CTL and A8-KO HCT116 cells. Mean ± SEM, 3 biological replicates, one-way ANOVA with Tukey’s multiple comparisons test. j Ribosomal elongation speed was measured using harringtonine chase. Polysome profiles of CTL cells or A8-KO cells treated with harringtonine (2 ug/mL) for indicated times. Monosome (80S) and polysomes were highlighted and the P/M ratio change was quantified in the right panel. k The polysome dissociation halftime in (g) was calculated. Mean ± SEM, 3 biological replicates, two-sided t-test. l The distributions of KRAS/ACTB mRNAs in the polysome fractions of (g). Mean ± SEM, 3 biological replicates, two-sided t-test. m The dissociation halftime of KRAS mRNA in (i) was calculated. Mean ± SEM, 3 biological replicates, two-sided t-test. Source data are provided as a Source Data file.