R-protein composition of budding yeast PTC deficient 80S ribosomes

Ribosomal RNA modifications are introduced during ribosome assembly by snoRNA and stand-alone modification enzyme based mechanisms in budding yeast. Lack of individual modifications in rRNA has a minor effect on yeast growth, ribosome biogenesis, and translation, which has complicated the definition of the biological function of rRNA modifications. We have constructed a S.cerevisiae strain lacking 7 25S rRNA modifications (Ψ2826, Um2921, Gm2619, m5C2870, Ψ2880, Um2724 and Gm2922)in peptidyl-transferase center. This strain exhibits severely compromised growth at 30°C, reduced level of global translation and actively translating ribosomes and cold sensitivity. Ribosomal protein composition of the mature 80S ribosomes was analyzed by SILAC based qMS approach. We have found that absence of PTC modifications around PTC does not affect significantly r-protein content of 80S ribosomes.