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Title90 pre-ribosome transformation into the primordial 40 subunit.
Journal, issue, pagesScience, Vol. 369, Issue 6510, Page 1470-1476, Year 2020
Publish dateSep 18, 2020
AuthorsJingdong Cheng / Benjamin Lau / Giuseppe La Venuta / Michael Ameismeier / Otto Berninghausen / Ed Hurt / Roland Beckmann /
PubMed AbstractProduction of small ribosomal subunits initially requires the formation of a 90 precursor followed by an enigmatic process of restructuring into the primordial pre-40 subunit. We elucidate this ...Production of small ribosomal subunits initially requires the formation of a 90 precursor followed by an enigmatic process of restructuring into the primordial pre-40 subunit. We elucidate this process by biochemical and cryo-electron microscopy analysis of intermediates along this pathway in yeast. First, the remodeling RNA helicase Dhr1 engages the 90 pre-ribosome, followed by Utp24 endonuclease-driven RNA cleavage at site A, thereby separating the 5'-external transcribed spacer (ETS) from 18 ribosomal RNA. Next, the 5'-ETS and 90 assembly factors become dislodged, but this occurs sequentially, not en bloc. Eventually, the primordial pre-40 emerges, still retaining some 90 factors including Dhr1, now ready to unwind the final small nucleolar U3-18 RNA hybrid. Our data shed light on the elusive 90 to pre-40 transition and clarify the principles of assembly and remodeling of large ribonucleoproteins.
External linksScience / PubMed:32943521
MethodsEM (single particle)
Resolution3.5 - 7.1 Å
Structure data

EMDB-11357, PDB-6zqa:
Cryo-EM structure of the 90S pre-ribosome from Saccharomyces cerevisiae, state A (Poly-Ala)
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-11358, PDB-6zqb:
Cryo-EM structure of the 90S pre-ribosome from Saccharomyces cerevisiae, state B2
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-11359, PDB-6zqc:
Cryo-EM structure of the 90S pre-ribosome from Saccharomyces cerevisiae, state Pre-A1
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-11360, PDB-6zqd:
Cryo-EM structure of the 90S pre-ribosome from Saccharomyces cerevisiae, state Post-A1
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-11361, PDB-6zqe:
Cryo-EM structure of the 90S pre-ribosome from Saccharomyces cerevisiae, state Dis-A (Poly-Ala)
Method: EM (single particle) / Resolution: 7.1 Å

EMDB-11362, PDB-6zqf:
Cryo-EM structure of the 90S pre-ribosome from Saccharomyces cerevisiae, state Dis-B (Poly-Ala)
Method: EM (single particle) / Resolution: 4.9 Å

EMDB-11363, PDB-6zqg:
Cryo-EM structure of the 90S pre-ribosome from Saccharomyces cerevisiae, state Dis-C
Method: EM (single particle) / Resolution: 3.5 Å

Chemicals

ChemComp-ZN:
ZINC ION / Zinc

ChemComp-MG:
MAGNESIUM ION / Magnesium

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

Source
  • saccharomyces cerevisiae (strain atcc 204508 / s288c) (yeast)
  • Baker's yeast (baker's yeast)
  • saccharomyces cerevisiae s288c (yeast)
  • accharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast)
KeywordsCryoelectron Microscopy / DEAD-box RNA Helicases / ECM16 protein, S cerevisiae / Nuclear Proteins / PNO1 protein, S cerevisiae / Protein Conformation / RNA Cleavage / RNA, Ribosomal, 18S / Ribosomal Proteins / Ribosome Subunits, Large, Eukaryotic / Ribosome Subunits, Small, Eukaryotic / Saccharomyces cerevisiae / Saccharomyces cerevisiae Proteins / Utp24 protein, S cerevisiae / RIBOSOME / 90S pre-ribosome / 40S pre-ribosome / A1 cleavage / Dhr1

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