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60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes

During the highly conserved process of eukaryotic ribosome formation, RNA follows a maturation path with well-defined, successive intermediates that dynamically associate with many pre-ribosomal proteins. A comprehensive description of the assembly process is still lacking. To obtain data on the tim...

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Autores principales: Lebreton, Alice, Rousselle, Jean-Claude, Lenormand, Pascal, Namane, Abdelkader, Jacquier, Alain, Fromont-Racine, Micheline, Saveanu, Cosmin
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2008
Materias:
RNA
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2528192/
https://www.ncbi.nlm.nih.gov/pubmed/18658244
http://dx.doi.org/10.1093/nar/gkn469
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author Lebreton, Alice
Rousselle, Jean-Claude
Lenormand, Pascal
Namane, Abdelkader
Jacquier, Alain
Fromont-Racine, Micheline
Saveanu, Cosmin
author_facet Lebreton, Alice
Rousselle, Jean-Claude
Lenormand, Pascal
Namane, Abdelkader
Jacquier, Alain
Fromont-Racine, Micheline
Saveanu, Cosmin
author_sort Lebreton, Alice
collection PubMed
description During the highly conserved process of eukaryotic ribosome formation, RNA follows a maturation path with well-defined, successive intermediates that dynamically associate with many pre-ribosomal proteins. A comprehensive description of the assembly process is still lacking. To obtain data on the timing and order of association of the different pre-ribosomal factors, a strategy consists in the use of pre-ribsomal particles isolated from mutants that block ribosome formation at different steps. Immunoblots, inherently limited to only a few factors, have been applied to evaluate the accumulation or decrease of pre-ribosomal intermediates under mutant conditions. For a global protein-level description of different 60S ribosomal subunit maturation intermediates in yeast, we have adapted a method of in vivo isotopic labelling and mass spectrometry to study pre-60S complexes isolated from strains in which rRNA processing was affected by individual depletion of five factors: Ebp2, Nog1, Nsa2, Nog2 or Pop3. We obtained quantitative data for 45 distinct pre-60S proteins and detected coordinated changes for over 30 pre-60S factors in the analysed mutants. These results led to the characterisation of the composition of early, intermediate and late pre-ribosomal complexes, specific for crucial maturation steps during 60S assembly in eukaryotes.
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spelling pubmed-25281922008-09-03 60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes Lebreton, Alice Rousselle, Jean-Claude Lenormand, Pascal Namane, Abdelkader Jacquier, Alain Fromont-Racine, Micheline Saveanu, Cosmin Nucleic Acids Res RNA During the highly conserved process of eukaryotic ribosome formation, RNA follows a maturation path with well-defined, successive intermediates that dynamically associate with many pre-ribosomal proteins. A comprehensive description of the assembly process is still lacking. To obtain data on the timing and order of association of the different pre-ribosomal factors, a strategy consists in the use of pre-ribsomal particles isolated from mutants that block ribosome formation at different steps. Immunoblots, inherently limited to only a few factors, have been applied to evaluate the accumulation or decrease of pre-ribosomal intermediates under mutant conditions. For a global protein-level description of different 60S ribosomal subunit maturation intermediates in yeast, we have adapted a method of in vivo isotopic labelling and mass spectrometry to study pre-60S complexes isolated from strains in which rRNA processing was affected by individual depletion of five factors: Ebp2, Nog1, Nsa2, Nog2 or Pop3. We obtained quantitative data for 45 distinct pre-60S proteins and detected coordinated changes for over 30 pre-60S factors in the analysed mutants. These results led to the characterisation of the composition of early, intermediate and late pre-ribosomal complexes, specific for crucial maturation steps during 60S assembly in eukaryotes. Oxford University Press 2008-09 2008-07-25 /pmc/articles/PMC2528192/ /pubmed/18658244 http://dx.doi.org/10.1093/nar/gkn469 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle RNA
Lebreton, Alice
Rousselle, Jean-Claude
Lenormand, Pascal
Namane, Abdelkader
Jacquier, Alain
Fromont-Racine, Micheline
Saveanu, Cosmin
60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes
title 60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes
title_full 60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes
title_fullStr 60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes
title_full_unstemmed 60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes
title_short 60S ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes
title_sort 60s ribosomal subunit assembly dynamics defined by semi-quantitative mass spectrometry of purified complexes
topic RNA
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2528192/
https://www.ncbi.nlm.nih.gov/pubmed/18658244
http://dx.doi.org/10.1093/nar/gkn469
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