Diversity of ribosomes at the level of rRNA variation associated with human health and disease.

With hundreds of copies of ribosomal DNA (rDNA) it is unknown whether they possess sequence variations that ultimately form different types of ribosomes. Here, we developed an algorithm for variant-calling between paralog genes (termed RGA) and compared rDNA variations with rRNA variations from long-read sequencing of translating ribosomes (RIBO-RT). Our analyses identified dozens of highly abundant rRNA variants, largely indels, that are incorporated into translationally active ribosomes and assemble into distinct ribosome subtypes encoded on different chromosomes. We developed an in-situ rRNA sequencing method (SWITCH-seq) revealing that variants are co-expressed within individual cells and found that they possess different structures. Lastly, we observed tissue-specific rRNA-subtype expression and linked specific rRNA variants to cancer. This study therefore reveals the variation landscape of translating ribosomes within human cells.