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TitleIn situ cryo-electron tomography reveals gradient organization of ribosome biogenesis in intact nucleoli.
Journal, issue, pagesNat Commun, Vol. 12, Issue 1, Page 5364, Year 2021
Publish dateSep 10, 2021
AuthorsPhilipp S Erdmann / Zhen Hou / Sven Klumpe / Sagar Khavnekar / Florian Beck / Florian Wilfling / Jürgen M Plitzko / Wolfgang Baumeister /
PubMed AbstractRibosomes comprise a large (LSU) and a small subunit (SSU) which are synthesized independently in the nucleolus before being exported into the cytoplasm, where they assemble into functional ribosomes. ...Ribosomes comprise a large (LSU) and a small subunit (SSU) which are synthesized independently in the nucleolus before being exported into the cytoplasm, where they assemble into functional ribosomes. Individual maturation steps have been analyzed in detail using biochemical methods, light microscopy and conventional electron microscopy (EM). In recent years, single particle analysis (SPA) has yielded molecular resolution structures of several pre-ribosomal intermediates. It falls short, however, of revealing the spatiotemporal sequence of ribosome biogenesis in the cellular context. Here, we present our study on native nucleoli in Chlamydomonas reinhardtii, in which we follow the formation of LSU and SSU precursors by in situ cryo-electron tomography (cryo-ET) and subtomogram averaging (STA). By combining both positional and molecular data, we reveal gradients of ribosome maturation within the granular component (GC), offering a new perspective on how the liquid-liquid-phase separation of the nucleolus supports ribosome biogenesis.
External linksNat Commun / PubMed:34508074 / PubMed Central
MethodsEM (subtomogram averaging) / EM (tomography)
Resolution23.6 - 34.7 Å
Structure data

EMDB-11043:
In situ cryo-electron tomography reveals layered organization of pre-ribosome maturation in nucleoli. Small Subunit Processome, Class 1
Method: EM (subtomogram averaging) / Resolution: 25.7 Å

EMDB-11044:
In situ cryo-electron tomography reveals layered organization of pre-ribosome maturation in nucleoli. Small Subunit Processome, Class 2
Method: EM (subtomogram averaging) / Resolution: 25.03 Å

EMDB-11045:
In situ cryo-electron tomography reveals layered organization of pre-ribosome maturation in nucleoli. Small Subunit Processome, Class 3
Method: EM (subtomogram averaging) / Resolution: 34.7 Å

EMDB-11046:
In situ cryo-electron tomography reveals layered organization of pre-ribosome maturation in nucleoli. Large Subunit Pre-Ribosome, Class 1
Method: EM (subtomogram averaging) / Resolution: 25.3 Å

EMDB-11047:
In situ cryo-electron tomography reveals layered organization of pre-ribosome maturation in nucleoli. Large Subunit Pre-Ribosome, Class 2
Method: EM (tomography)

EMDB-11048:
In situ cryo-electron tomography reveals layered organization of pre-ribosome maturation in nucleoli. Large Subunit Pre-Ribosome, Class 3
Method: EM (subtomogram averaging) / Resolution: 23.6 Å

Source
  • Chlamydomonas reinhardtii (plant)

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