Structure and assembly model for the Trypanosoma cruzi 60S ribosomal subunit

Significance The pathogenic trypanosomatids—Trypanosoma cruzi, Trypanosoma brucei, and Leishmania spp.—are the causative agents of Chagas disease, African trypanosomiasis, and leishmaniasis, respectively. These diseases, with high morbidity and mortality rates, affect millions of people worldwide. Current treatments typically use drugs with high toxicity and marginal efficacy. Here we present, a 2.5-Å structure of the T. cruzi ribosome large subunit by single-particle cryo-EM. Our structure highlights distinctive trypanosome interactions and has allowed us to propose a tentative model for assembly of the 60S large ribosomal subunit. These atomic details highlighting trypanosome-specific interactions and the differences between T. cruzi and the human ribosome can be used directly for structure-based drug design of antitrypanosome drugs. Ribosomes of trypanosomatids, a family of protozoan parasites causing debilitating human diseases, possess multiply fragmented rRNAs that together are analogous to 28S rRNA, unusually large rRNA expansion segments, and r-protein variations compared with other eukaryotic ribosomes. To investigate the architecture of the trypanosomatid ribosomes, we determined the 2.5-Å structure of the Trypanosoma cruzi ribosome large subunit by single-particle cryo-EM. Examination of this structure and comparative analysis of the yeast ribosomal assembly pathway allowed us to develop a stepwise assembly model for the eight pieces of the large subunit rRNAs and a number of ancillary “glue” proteins. This model can be applied to the characterization of Trypanosoma brucei and Leishmania spp. ribosomes as well. Together with other details, our atomic-level structure may provide a foundation for structure-based design of antitrypanosome drugs.

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