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TitleNear-physiological in vitro assembly of 50S ribosomes involves parallel pathways.
Journal, issue, pagesNucleic Acids Res, Vol. 51, Issue 6, Page 2862-2876, Year 2023
Publish dateApr 11, 2023
AuthorsXiyu Dong / Lili K Doerfel / Kai Sheng / Jessica N Rabuck-Gibbons / Anna M Popova / Dmitry Lyumkis / James R Williamson /
PubMed AbstractUnderstanding 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 linksNucleic Acids Res / PubMed:36864669 / PubMed Central
MethodsEM (single particle)
Resolution4.5 - 8.8 Å
Structure data

EMDB-29042: Structure of iSAT large ribosomal subunit assembly intermediate - Class B2
Method: EM (single particle) / Resolution: 6.0 Å

EMDB-29056: Structure of iSAT large ribosomal subunit assembly intermediate - Class B1
Method: EM (single particle) / Resolution: 7.0 Å

EMDB-29057: Structure of iSAT large ribosomal subunit assembly intermediate - Class G1
Method: EM (single particle) / Resolution: 8.8 Å

EMDB-29058: Structure of iSAT large ribosomal subunit assembly intermediate - Class C1
Method: EM (single particle) / Resolution: 6.5 Å

EMDB-29059: Structure of iSAT large ribosomal subunit assembly intermediate - Class C2
Method: EM (single particle) / Resolution: 4.5 Å

EMDB-29060: Structure of iSAT large ribosomal subunit assembly intermediate - Class C3
Method: EM (single particle) / Resolution: 5.2 Å

EMDB-29061: Structure of iSAT large ribosomal subunit assembly intermediate - Class C4
Method: EM (single particle) / Resolution: 6.6 Å

EMDB-29062: Structure of iSAT large ribosomal subunit assembly intermediate - Class E1
Method: EM (single particle) / Resolution: 6.6 Å

EMDB-29063: Structure of iSAT large ribosomal subunit assembly intermediate - Class E2
Method: EM (single particle) / Resolution: 5.8 Å

EMDB-29064: Structure of iSAT large ribosomal subunit assembly intermediate - Class E3
Method: EM (single particle) / Resolution: 5.1 Å

EMDB-29065: Structure of iSAT large ribosomal subunit assembly intermediate - Class E4
Method: EM (single particle) / Resolution: 5.7 Å

EMDB-29066: Structure of iSAT large ribosomal subunit assembly intermediate - Class E5
Method: EM (single particle) / Resolution: 5.2 Å

EMDB-29067: Structure of iSAT large ribosomal subunit assembly intermediate - Class E6
Method: EM (single particle) / Resolution: 4.8 Å

Source
  • Escherichia coli (E. coli)

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