Mechanism of completion of peptidyltransferase centre assembly in eukaryotes

DOI: 10.7554/eLife.44904

Authors: Vasileios Kargas (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Pablo Castro-Hartmann (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Norberto Escudero-Urquijo (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Kyle Dent (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Christine Hilcenko (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Carolin Sailer (University of Konstanz) , Gertrude Zisser (University of Graz) , Maria J. Marques-Carvalho (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Simone Pellegrino (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Leszek Wawiórka (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge; Maria Curie-Skłodowska University) , Stefan M. V. Freund (MRC Laboratory of Molecular Biology) , Jane L. Wagstaff (MRC Laboratory of Molecular Biology) , Antonina Andreeva (MRC Laboratory of Molecular Biology) , Alexandre Faille (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge) , Edwin Chen (University of Leeds) , Florian Stengel (University of Konstanz) , Helmut Bergler (University of Graz) , Alan John Warren (Cambridge Institute for Medical Research; Wellcome Trust–Medical Research Council Stem Cell Institute, University of Cambridge)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Elife , VOL 8

State: Published (Approved)
Published: May 2019
Diamond Proposal Number(s): 14359 , 15368 , 16940 , 17057 , 18078 , 18328

Abstract: During their final maturation in the cytoplasm, pre-60S ribosomal particles are converted to translation-competent large ribosomal subunits. Here, we present the mechanism of peptidyltransferase centre (PTC) completion that explains how integration of the last ribosomal proteins is coupled to release of the nuclear export adaptor Nmd3. Single-particle cryo-EM reveals that eL40 recruitment stabilises helix 89 to form the uL16 binding site. The loading of uL16 unhooks helix 38 from Nmd3 to adopt its mature conformation. In turn, partial retraction of the L1 stalk is coupled to a conformational switch in Nmd3 that allows the uL16 P-site loop to fully accommodate into the PTC where it competes with Nmd3 for an overlapping binding site (base A2971). Our data reveal how the central functional site of the ribosome is sculpted and suggest how the formation of translation-competent 60S subunits is disrupted in leukaemia-associated ribosomopathies.

Journal Keywords: S. cerevisiae; SBDS; Shwachman-Diamond syndrome; cryo-EM; leukaemia; molecular biophysics; ribosome; ribosomopathy; structural biology

Diamond Keywords: Leukaemia

Subject Areas: Biology and Bio-materials, Medicine

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios II-Titan Krios II at Diamond

Added On: 30/09/2019 11:06

Documents:
elife-44904-v2.pdf

Discipline Tags:

Non-Communicable Diseases Health & Wellbeing Cancer Structural biology Life Sciences & Biotech

Technical Tags:

Microscopy Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)