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Title | Near-physiological in vitro assembly of 50S ribosomes involves parallel pathways. |
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Journal, issue, pages | Nucleic Acids Res, Vol. 51, Issue 6, Page 2862-2876, Year 2023 |
Publish date | Apr 11, 2023 |
Authors | Xiyu Dong / Lili K Doerfel / Kai Sheng / Jessica N Rabuck-Gibbons / Anna M Popova / Dmitry Lyumkis / James R Williamson / |
PubMed Abstract | Understanding the assembly principles of biological macromolecular complexes remains a significant challenge, due to the complexity of the systems and the difficulties in developing experimental ...Understanding the assembly principles of biological macromolecular complexes remains a significant challenge, due to the complexity of the systems and the difficulties in developing experimental approaches. As a ribonucleoprotein complex, the ribosome serves as a model system for the profiling of macromolecular complex assembly. In this work, we report an ensemble of large ribosomal subunit intermediate structures that accumulate during synthesis in a near-physiological and co-transcriptional in vitro reconstitution system. Thirteen pre-50S intermediate maps covering the entire assembly process were resolved using cryo-EM single-particle analysis and heterogeneous subclassification. Segmentation of the set of density maps reveals that the 50S ribosome intermediates assemble based on fourteen cooperative assembly blocks, including the smallest assembly core reported to date, which is composed of a 600-nucleotide-long folded rRNA and three ribosomal proteins. The cooperative blocks assemble onto the assembly core following defined dependencies, revealing the parallel pathways at both early and late assembly stages of the 50S subunit. |
External links | Nucleic Acids Res / PubMed:36864669 / PubMed Central |
Methods | EM (single particle) |
Resolution | 4.5 - 8.8 Å |
Structure data | EMDB-29042: Structure of iSAT large ribosomal subunit assembly intermediate - Class B2 EMDB-29056: Structure of iSAT large ribosomal subunit assembly intermediate - Class B1 EMDB-29057: Structure of iSAT large ribosomal subunit assembly intermediate - Class G1 EMDB-29058: Structure of iSAT large ribosomal subunit assembly intermediate - Class C1 EMDB-29059: Structure of iSAT large ribosomal subunit assembly intermediate - Class C2 EMDB-29060: Structure of iSAT large ribosomal subunit assembly intermediate - Class C3 EMDB-29061: Structure of iSAT large ribosomal subunit assembly intermediate - Class C4 EMDB-29062: Structure of iSAT large ribosomal subunit assembly intermediate - Class E1 EMDB-29063: Structure of iSAT large ribosomal subunit assembly intermediate - Class E2 EMDB-29064: Structure of iSAT large ribosomal subunit assembly intermediate - Class E3 EMDB-29065: Structure of iSAT large ribosomal subunit assembly intermediate - Class E4 EMDB-29066: Structure of iSAT large ribosomal subunit assembly intermediate - Class E5 EMDB-29067: Structure of iSAT large ribosomal subunit assembly intermediate - Class E6 |
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