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TitleCoupling of 5S RNP rotation with maturation of functional centers during large ribosomal subunit assembly.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 3751, Year 2020
Publish dateJul 27, 2020
AuthorsJelena Micic / Yu Li / Shan Wu / Daniel Wilson / Beril Tutuncuoglu / Ning Gao / John L Woolford /
PubMed AbstractThe protein composition and structure of assembling 60S ribosomal subunits undergo numerous changes as pre-ribosomes transition from the nucleolus to the nucleoplasm. This includes stable anchoring ...The protein composition and structure of assembling 60S ribosomal subunits undergo numerous changes as pre-ribosomes transition from the nucleolus to the nucleoplasm. This includes stable anchoring of the Rpf2 subcomplex containing 5S rRNA, rpL5, rpL11, Rpf2 and Rrs1, which initially docks onto the flexible domain V of rRNA at earlier stages of assembly. In this work, we tested the function of the C-terminal domain (CTD) of Rpf2 during these anchoring steps, by truncating this extension and assaying effects on middle stages of subunit maturation. The rpf2Δ255-344 mutation affects proper folding of rRNA helices H68-70 during anchoring of the Rpf2 subcomplex. In addition, several assembly factors (AFs) are absent from pre-ribosomes or in altered conformations. Consequently, major remodeling events fail to occur: rotation of the 5S RNP, maturation of the peptidyl transferase center (PTC) and the nascent polypeptide exit tunnel (NPET), and export of assembling subunits to the cytoplasm.
External linksNat Commun / PubMed:32719344 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 6.0 Å
Structure data

30108

6m62

EMDB-30108, PDB-6m62:
Cryo-Em structure of eukaryotic pre-60S ribosome subunit from Saccharomyces cerevisiae rpf2 delta 255-344 strain, C4 state.
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-30109:
Eukaryotic pre-60S ribosomal subunits from Saccharomyces cerevisiae Sda1 depletion strain, E2 state
Method: EM (single particle) / Resolution: 4.8 Å

EMDB-30110:
Eukaryotic pre-60S ribosomal subunit from Saccharomyces cerevisiae rpf2delta255-344 strain, C1 state at 6 Angstroms resolution.
Method: EM (single particle) / Resolution: 6.0 Å

EMDB-30111:
Eukaryotic pre-60S ribosomal subunit from Saccharomyces cerevisiae rpf2delta255-344 strain, C2 state at 5.9 Angstroms resolution
Method: EM (single particle) / Resolution: 5.9 Å

EMDB-30112:
Cryo-EM structure of pre-60S ribosomal subunit from Saccharomyces cerevisiae rpf2 delta 255-344 strain, C3 state at 3.9 Angstroms resolution.
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-30113:
Eukaryotic pre-60S ribosomal subunits from Saccharomyces cerevisiae Sda1 depletion strain, E1 state
Method: EM (single particle) / Resolution: 5.9 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

Source
  • saccharomyces cerevisiae s288c (yeast)
KeywordsRIBOSOME / pre-60s / Rpf2

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