PXD019058

PXD019058 is an original dataset announced via ProteomeXchange.

Title	Ribosome protein composition in wt and 23S+5S rRNA ribosomes
Description	The 120-nt long 5S rRNA, is an indispensable component of cytoplasmic ribosomes in all living organisms. The functions of 5S rRNA and the reasons for its evolutionary preservation as an independent molecule remain unclear. Here we used ribosome engineering to investigate whether maintaining 5S rRNA as an independent molecule is critical for ribosome function and cell survival. By fusing circularly permutated 5S rRNA (cp5S) with 23S rRNA and deleting all wild type 5S rRNA genes, we generated an Escherichia coli strain completely devoid of free 5S rRNA. Viability of the engineered cells demonstrates that autonomous 5S rRNA is not required for cell growth at 37°C and is unlikely to have essential functions outside the ribosome. The fully-assembled ribosomes carrying 23S-cp5S hybrid rRNA and lacking free 5S rRNA are highly active in translation. However, the engineered cells accumulate aberrant 50S subunits that are unable to form stable 70S ribosomes. Cryo-EM analysis revealed a dramatically malformed peptidyl transferase center in the misassembled 50S subunits. The results of our experiments argue that the key evolutionary force preserving the autonomous nature of the smallest rRNA is its role in ribosome biogenesis.
HostingRepository	PRIDE
AnnounceDate	2020-06-18
AnnouncementXML	Submission_2020-06-17_23:32:47.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD019058
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Kaspar Reier
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	Label:2H(4); Oxidation; Label:13C(6); monohydroxylated residue; Carbamidomethyl; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap XL

Revision	Datetime	Status	ChangeLog Entry
0	2020-05-08 05:49:33	ID requested
⏵ 1	2020-06-17 23:32:48	announced

Huang S, Aleksashin NA, Loveland AB, Klepacki D, Reier K, Kefi A, Szal T, Remme J, Jaeger L, V, á, zquez-Laslop N, Korostelev AA, Mankin AS, Ribosome engineering reveals the importance of 5S rRNA autonomy for ribosome assembly. Nat Commun, 11(1):2900(2020) [pubmed]

submitter keyword: Ribosome, Mass-spectrometry, proteins

Tiina Tamm
contact affiliation	Institute of molecular and cellular biology, University of Tartu, Estonia
contact email	tiina.tamm.001@ut.ee
lab head
Kaspar Reier
contact affiliation	University of Tatu
contact email	kaspar.reier@ut.ee
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD019058
     1. Label: PRIDE project
     2. Name: Ribosome protein composition in wt and 23S+5S rRNA ribosomes