The dynamic rRNA ribomethylome drives stemness in acute myeloid leukemia [mRNA_FBL_KD]

Eukaryotic ribosomal RNA carries diverse posttranscriptional modifications, the function of which are mostly unexplored. The evolutionarily conserved 2'-O-methylation (2'-O-Me) occurs at more than 100 sites and is essential for ribosome biogenesis. Plasticity of 2´-O-Me in ribosomes and its functional consequences in human disease remain to be elucidated. The effect of rRNA 2'-O-Me on protein translation was investigated by nascent proteomcis analysis coupled with RNA-Seq in human leukemia cell line Kasumi-1 with and without knockdown of 2´-O-methyltransferase FBL. Overall design: Human AML cell line expressing doxycycline inducible control shRNA (shLUC) or FBL specific shRNA (shFBL) was used. The cells were treated with 100 ng/ml doxycycline for 72 hours for FBL knockown. The newly newly synthesized proteins and total RNA were isolated from the control and FBL knockdown cells. Total RNA was submitted to RNA-Seq and nascent proteins were used for mass spectrometry proteomics. The changes in nascent proteins were normalized by mRNA alteration.