Pseudouridine increases ribosome stability in a thermophilic eukaryote

RNA modifications alter stability, folding space and interaction network of RNA molecules. Ribosomal RNA (rRNA) modifications stabilize the structure of ribosomes and are critical for their function. They are known to cluster around functionally important sites such as the peptidyl transferase center, ribosomal subunit bridges and the polypeptide tunnel. Here, we investigate the rRNA modifications of the thermophilic fungus Chaetomium thermophilum (ct), a unique tool and model system for eukaryotic structural stability and thermophily. Using LC-MS/MS, orthogonal second and third generation RNA-sequencing and high-resolution cryo-electron microscopy (cryo-EM), we describe a cross-correlating method to assign and quantify all ct rRNA modifications.