Cryo-EM captures early ribosome assembly in action.


Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
17 02 2023
Historique:
received: 10 11 2022
accepted: 08 02 2023
entrez: 16 2 2023
pubmed: 17 2 2023
medline: 22 2 2023
Statut: epublish

Résumé

Ribosome biogenesis is a fundamental multi-step cellular process in all domains of life that involves the production, processing, folding, and modification of ribosomal RNAs (rRNAs) and ribosomal proteins. To obtain insights into the still unexplored early assembly phase of the bacterial 50S subunit, we exploited a minimal in vitro reconstitution system using purified ribosomal components and scalable reaction conditions. Time-limited assembly assays combined with cryo-EM analysis visualizes the structurally complex assembly pathway starting with a particle consisting of ordered density for only ~500 nucleotides of 23S rRNA domain I and three ribosomal proteins. In addition, our structural analysis reveals that early 50S assembly occurs in a domain-wise fashion, while late 50S assembly proceeds incrementally. Furthermore, we find that both ribosomal proteins and folded rRNA helices, occupying surface exposed regions on pre-50S particles, induce, or stabilize rRNA folds within adjacent regions, thereby creating cooperativity.

Identifiants

pubmed: 36797249
doi: 10.1038/s41467-023-36607-9
pii: 10.1038/s41467-023-36607-9
pmc: PMC9935924
doi:

Substances chimiques

Ribosomal Proteins 0
RNA, Ribosomal, 23S 0
Nucleotides 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

898

Informations de copyright

© 2023. The Author(s).

Références

RNA. 2008 Oct;14(10):1999-2012
pubmed: 18755834
Nat Commun. 2021 Jun 16;12(1):3673
pubmed: 34135318
Mol Cell. 2021 Mar 18;81(6):1200-1215.e9
pubmed: 33639093
Cell. 2001 Nov 30;107(5):679-88
pubmed: 11733066
Science. 1989 May 19;244(4906):783-90
pubmed: 2658053
Proc Natl Acad Sci U S A. 1968 Mar;59(3):777-84
pubmed: 4868216
PLoS Biol. 2014 May 20;12(5):e1001866
pubmed: 24844575
Proc Natl Acad Sci U S A. 2020 Jul 7;117(27):15609-15619
pubmed: 32571953
Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11681-5
pubmed: 8524828
Elife. 2018 Nov 09;7:
pubmed: 30412051
Nat Methods. 2013 Jan;10(1):74-6
pubmed: 23202432
Mol Cell. 2020 Aug 20;79(4):629-644.e4
pubmed: 32679035
Elife. 2019 May 22;8:
pubmed: 31115337
Cell. 2016 Dec 1;167(6):1610-1622.e15
pubmed: 27912064
Trends Biochem Sci. 2021 Nov;46(11):889-901
pubmed: 34176739
Nature. 2016 May 25;534(7605):133-7
pubmed: 27251291
Nat Commun. 2021 Jun 16;12(1):3671
pubmed: 34135320
J Struct Biol. 2016 Jan;193(1):1-12
pubmed: 26592709
Nat Methods. 2017 Mar;14(3):290-296
pubmed: 28165473
Cell. 2019 Nov 27;179(6):1370-1381.e12
pubmed: 31761536
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4713-7
pubmed: 4612527
Microbiol Mol Biol Rev. 2007 Sep;71(3):477-94
pubmed: 17804668
Nat Commun. 2021 Jul 27;12(1):4544
pubmed: 34315873
Nat Commun. 2021 Jun 16;12(1):3672
pubmed: 34135319
Cell. 2019 Nov 27;179(6):1357-1369.e16
pubmed: 31761533
Nucleic Acids Res. 2013 Aug;41(14):7073-83
pubmed: 23700310
Nat Commun. 2019 Feb 27;10(1):958
pubmed: 30814529
Proc Natl Acad Sci U S A. 1984 Jul;81(14):4290-3
pubmed: 6379642
Mol Cell. 2018 Jun 7;70(5):881-893.e3
pubmed: 29883607
Annu Rev Biochem. 2011;80:501-26
pubmed: 21529161
J Biol Chem. 1987 Jun 25;262(18):8826-33
pubmed: 3298242
J Struct Biol. 2013 Nov;184(2):321-8
pubmed: 24075951
Biochimie. 1991 Jun;73(6):739-55
pubmed: 1764520
Cell. 2017 Dec 14;171(7):1599-1610.e14
pubmed: 29245012
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
Science. 2000 Aug 11;289(5481):905-20
pubmed: 10937989
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
pubmed: 20124702
Mol Cell. 2020 Aug 20;79(4):615-628.e5
pubmed: 32668200
Nucleic Acids Res. 2014 Mar;42(5):3419-35
pubmed: 24335279
Elife. 2020 Sep 14;9:
pubmed: 32924932
J Struct Biol. 1996 Jan-Feb;116(1):190-9
pubmed: 8742743
Nucleic Acids Res. 2016 Sep 30;44(17):8442-55
pubmed: 27484475
Nucleic Acids Res. 2014 Dec 1;42(21):13430-9
pubmed: 25389271
Elife. 2021 Oct 05;10:
pubmed: 34609277
Nucleic Acids Res. 1981 Nov 25;9(22):6167-89
pubmed: 7031608
J Mol Biol. 1976 Nov 15;107(4):585-99
pubmed: 794489
Nature. 2018 Apr 5;556(7699):126-129
pubmed: 29512650
Proc Natl Acad Sci U S A. 2015 Aug 25;112(34):E4707-16
pubmed: 26261349
Protein Sci. 2018 Jan;27(1):14-25
pubmed: 28710774

Auteurs

Bo Qin (B)

Institut für Medizinische Physik und Biophysik, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin, Berlin, Germany.

Simon M Lauer (SM)

Institut für Medizinische Physik und Biophysik, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin, Berlin, Germany.

Annika Balke (A)

Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Institute of Medical Physics and Biophysics, Group Protein X-ray Crystallography and Signal Transduction, Charitéplatz 1, D-10117, Berlin, Germany.

Carlos H Vieira-Vieira (CH)

Proteome Dynamics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Robert-Rössle-Str. 10, 13125, Berlin, Germany.
Faculty of Life Sciences, Humboldt Universität zu Berlin, Berlin, Germany.

Jörg Bürger (J)

Institut für Medizinische Physik und Biophysik, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin, Berlin, Germany.
Microscopy and Cryo-Electron Microscopy Service Group, Max Planck Institute for Molecular Genetics, Ihnestr. 63-73, 14195, Berlin, Germany.

Thorsten Mielke (T)

Microscopy and Cryo-Electron Microscopy Service Group, Max Planck Institute for Molecular Genetics, Ihnestr. 63-73, 14195, Berlin, Germany.

Matthias Selbach (M)

Proteome Dynamics, Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Robert-Rössle-Str. 10, 13125, Berlin, Germany.
Charité -Universitätsmedizin Berlin, 10117, Berlin, Germany.

Patrick Scheerer (P)

Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Institute of Medical Physics and Biophysics, Group Protein X-ray Crystallography and Signal Transduction, Charitéplatz 1, D-10117, Berlin, Germany.

Christian M T Spahn (CMT)

Institut für Medizinische Physik und Biophysik, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin, Berlin, Germany. christian.spahn@charite.de.

Rainer Nikolay (R)

Institut für Medizinische Physik und Biophysik, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin, Berlin, Germany. nikolay@molgen.mpg.de.
Department of Genome Regulation, Max Planck Institute for Molecular Genetics, Ihnestr. 63-73, 14195, Berlin, Germany. nikolay@molgen.mpg.de.

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Classifications MeSH