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TitleIn vitro structural maturation of an early stage pre-40S particle coupled with U3 snoRNA release and central pseudoknot formation.
Journal, issue, pagesNucleic Acids Res, Vol. 50, Issue 20, Page 11916-11923, Year 2022
Publish dateNov 11, 2022
AuthorsJingdong Cheng / Giuseppe La Venuta / Benjamin Lau / Otto Berninghausen / Roland Beckmann / Ed Hurt /
PubMed AbstractThe transition of the 90S to the pre-40S pre-ribosome is a decisive step in eukaryotic small subunit biogenesis leading to a first pre-40S intermediate (state Dis-C or primordial pre-40S), where the ...The transition of the 90S to the pre-40S pre-ribosome is a decisive step in eukaryotic small subunit biogenesis leading to a first pre-40S intermediate (state Dis-C or primordial pre-40S), where the U3 snoRNA keeps the nascent 18S rRNA locally immature. We in vitro reconstitute the ATP-dependent U3 release from this particle, catalyzed by the helicase Dhr1, and follow this process by cryo-EM revealing two successive pre-40S intermediates, Dis-D and Dis-E. The latter has lost not only U3 but all residual 90S factors including the GTPase Bms1. In vitro remodeling likewise induced the formation of the central pseudoknot, a universally conserved tertiary RNA structure that comprises the core of the small subunit decoding center. Thus, we could structurally reveal a key tertiary RNA folding step that is essential to form the active 40S subunit.
External linksNucleic Acids Res / PubMed:36263816 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 3.5 Å
Structure data

32790

7wtl

EMDB-32790, PDB-7wtl:
Cryo-EM structure of a yeast pre-40S ribosomal subunit - State Dis-D
Method: EM (single particle) / Resolution: 3.3 Å

32791

7wtm

EMDB-32791, PDB-7wtm:
Cryo-EM structure of a yeast pre-40S ribosomal subunit - State Dis-E
Method: EM (single particle) / Resolution: 3.5 Å

Chemicals

ChemComp-ZN:
Unknown entry

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

ChemComp-MG:
Unknown entry

Source
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsRIBOSOME / ribosome biogenesis / 40S ribosome

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