Good Vibrations: Structural Remodeling of Maturing Yeast Pre-40S Ribosomal Particles Followed by Cryo-Electron Microscopy

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Ramtin ShayanCélia Plisson-Chastang

Abstract

Assembly of eukaryotic ribosomal subunits is a very complex and sequential process that starts in the nucleolus and finishes in the cytoplasm with the formation of functional ribosomes. Over the past few years, characterization of the many molecular events underlying eukaryotic ribosome biogenesis has been drastically improved by the "resolution revolution" of cryo-electron microscopy (cryo-EM). However, if very early maturation events have been well characterized for both yeast ribosomal subunits, little is known regarding the final maturation steps occurring to the small (40S) ribosomal subunit. To try to bridge this gap, we have used proteomics together with cryo-EM and single particle analysis to characterize yeast pre-40S particles containing the ribosome biogenesis factor Tsr1. Our analyses lead us to refine the timing of the early pre-40S particle maturation steps. Furthermore, we suggest that after an early and structurally stable stage, the beak and platform domains of pre-40S particles enter a "vibrating" or "wriggling" stage, that might be involved in the final maturation of 18S rRNA as well as the fitting of late ribosomal proteins into their mature position.

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Citations

Oct 25, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Quentin Vicens
Nov 3, 2020·Nucleic Acids Research·Lisbeth-Carolina AguilarBen Montpetit
May 9, 2021·Nucleic Acids Research·Guillaume PosticPierre Tufféry

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Methods Mentioned

BETA
tandem affinity purification
affinity purification
electrophoresis
acetylation

Software Mentioned

XCalibur
Mascot
ProSMART
UCSF Chimera
REFMAC5
RELION
Proline
Coot
Phenix
MOTIONCORR2

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