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TitleStructures of the eukaryotic ribosome and its translational states in situ.
Journal, issue, pagesNat Commun, Vol. 13, Issue 1, Page 7435, Year 2022
Publish dateDec 2, 2022
AuthorsPatrick C Hoffmann / Jan Philipp Kreysing / Iskander Khusainov / Maarten W Tuijtel / Sonja Welsch / Martin Beck /
PubMed AbstractRibosomes translate genetic information into primary structure. During translation, various cofactors transiently bind to the ribosome that undergoes prominent conformational and structural changes. ...Ribosomes translate genetic information into primary structure. During translation, various cofactors transiently bind to the ribosome that undergoes prominent conformational and structural changes. Different translational states of ribosomes have been well characterized in vitro. However, to which extent the known translational states are representative of the native situation inside cells has thus far only been addressed in prokaryotes. Here, we apply cryo-electron tomography to cryo-FIB milled Dictyostelium discoideum cells combined with subtomogram averaging and classification. We obtain an in situ structure that is locally resolved up to 3 Angstrom, the distribution of eukaryotic ribosome translational states, and unique arrangement of rRNA expansion segments. Our work demonstrates the use of in situ structural biology techniques for identifying distinct ribosome states within the cellular environment.
External linksNat Commun / PubMed:36460643 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution3.8 - 16.0 Å
Structure data

EMDB-15807: In situ Dictyostelium discoideum 80S ribosome from dataset2 (1.223A/px)
Method: EM (subtomogram averaging) / Resolution: 3.8 Å

EMDB-15808: In situ Dictyostelium discoideum 80S ribosome from dataset1 (2.176A/px)
Method: EM (subtomogram averaging) / Resolution: 4.5 Å

EMDB-15809: In situ Dictyostelium discoideum membrane-bound 80S ribosome
Method: EM (subtomogram averaging) / Resolution: 10.3 Å

EMDB-15810: In situ Dictyostelium discoideum 80S ribosome with eEF1A, A/T-tRNA and P-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 6.9 Å

EMDB-15811: In situ Dictyostelium discoideum 80S ribosome with eEF1A, A/T-tRNA, P-tRNA and E-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 16.0 Å

EMDB-15812: In situ Dictyostelium discoideum 80S ribosome with A-tRNA and P-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 7.6 Å

EMDB-15813: In situ Dictyostelium discoideum 80S ribosome with P-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 10.8 Å

EMDB-15814: In situ Dictyostelium discoideum 80S ribosome with factor, A-tRNA and P-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 10.0 Å

EMDB-15815: In situ Dictyostelium discoideum 80S ribosome with eEF2, aa/P-tRNA and P/E-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 9.8 Å

EMDB-15816: In situ Dictyostelium discoideum 80S ribosome with eEF2, ap/P-tRNA and P/E-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 11.6 Å

EMDB-15843: In situ Dictyostelium discoideum 80S ribosome with eEF2 bound (substate1)
Method: EM (subtomogram averaging) / Resolution: 8.6 Å

EMDB-15844: In situ Dictyostelium discoideum 80S ribosome with eEF2 bound (substate2)
Method: EM (subtomogram averaging) / Resolution: 12.0 Å

EMDB-15845: In situ Dictyostelium discoideum 80S ribosome with eEF2, P-tRNA, and E-tRNA bound
Method: EM (subtomogram averaging) / Resolution: 11.0 Å

Source
  • Dictyostelium discoideum AX2 (eukaryote)

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