PXD030312

PXD030312 is an original dataset announced via ProteomeXchange.

Title	Visualization of early ribosome assembly in action
Description	Ribosomes, macromolecular machines producing a cell´s protein content, are formed from their RNA and protein components in a dynamic process referred to as ribosome assembly. While ribosome assembly in the cell starts with the successive synthesis of ribosomal RNA (rRNA) by the RNA polymerase holoenzyme, and requires numerous assembly factors, (Kaczanowska & Ryden-Aulin, 2007; Shajani et al., 2011), the process can be accomplished in vitro, using purified ribosomal components and scalable reaction conditions (Nierhaus & Dohme, 1974; Traub & Nomura, 1968). To obtain structural and conceptual insights in the early phase of the process, we performed the in vitro assembly reaction of the bacterial 50S subunit as a time course reaction, analyzed samples by sucrose density gradient ultracentrifugation, activity assay, quantitative mass spectrometry (qMS) and cryo-electron microscopy (cryo-EM). Our structural analysis reveals that early 50 assembly initiates with 23S rRNA domain I and occurs in a domain-wise fashion, while late 50S assembly proceeds incrementally. Notably, in both phases parallel pathways are utilized. 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. Our study provides new insights into fundamental principles governing ribosome assembly.
HostingRepository	PRIDE
AnnounceDate	2022-09-05
AnnouncementXML	Submission_2022-09-05_04:20:27.770.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Carlos Vieira
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	carbamoylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-12-11 15:31:19	ID requested
⏵ 1	2022-09-05 04:20:28	announced

Nikolay R, Hilal T, Schmidt S, Qin B, Schwefel D, Vieira-Vieira CH, Mielke T, B, ü, rger J, Loerke J, Amikura K, Fl, ü, gel T, Ueda T, Selbach M, Deuerling E, Spahn CMT, Snapshots of native pre-50S ribosomes reveal a biogenesis factor network and evolutionary specialization. Mol Cell, 81(6):1200-1215.e9(2021) [pubmed]

submitter keyword: Ribosome assembly

Matthias Selbach
contact affiliation	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
contact email	matthias.selbach@mdc-berlin.de
lab head
Carlos Vieira
contact affiliation	Max-Delbrück-Centrum für Molekulare Medizin in der Helmholtz-Gemeinschaft
contact email	carlos.vieira@mdc-berlin.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2022/09/PXD030312

PRIDE project URI

• PRIDE
  1. PXD030312
     1. Label: PRIDE project
     2. Name: Visualization of early ribosome assembly in action