Ribosome engineering reveals the importance of 5S rRNA autonomy for ribosome assembly.
Catalytic Domain
Cryoelectron Microscopy
DNA-Binding Proteins
/ metabolism
Escherichia coli
/ metabolism
Escherichia coli Proteins
/ metabolism
Gene Expression Regulation
Genetic Engineering
Mutation
Nucleic Acid Conformation
Peptidyl Transferases
/ metabolism
RNA, Bacterial
RNA, Ribosomal, 23S
/ metabolism
RNA, Ribosomal, 5S
/ metabolism
Rec A Recombinases
/ metabolism
Ribosomal Proteins
/ metabolism
Ribosome Subunits, Large, Bacterial
/ metabolism
Ribosomes
/ metabolism
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
09 06 2020
09 06 2020
Historique:
received:
12
03
2020
accepted:
11
05
2020
entrez:
11
6
2020
pubmed:
11
6
2020
medline:
28
8
2020
Statut:
epublish
Résumé
5S rRNA is an indispensable component of cytoplasmic ribosomes in all species. 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 5S rRNA autonomy is critical for ribosome function and cell survival. By linking circularly permutated 5S rRNA with 23S rRNA we generated a bacterial strain devoid of free 5S rRNA. Viability of the engineered cells demonstrates that autonomous 5S rRNA is dispensable for cell growth under standard conditions and is unlikely to have essential functions outside the ribosome. The fully assembled ribosomes carrying 23S-5S rRNA are highly active in translation. However, the engineered cells accumulate aberrant 50S subunits unable to form stable 70S ribosomes. Cryo-EM analysis revealed a malformed peptidyl transferase center in the misassembled 50S subunits. Our results argue that the autonomy of 5S rRNA is preserved due to its role in ribosome biogenesis.
Identifiants
pubmed: 32518240
doi: 10.1038/s41467-020-16694-8
pii: 10.1038/s41467-020-16694-8
pmc: PMC7283268
doi:
Substances chimiques
DNA-Binding Proteins
0
Escherichia coli Proteins
0
RNA, Bacterial
0
RNA, Ribosomal, 23S
0
RNA, Ribosomal, 5S
0
Ribosomal Proteins
0
recA protein, E coli
0
Peptidyl Transferases
EC 2.3.2.12
Rec A Recombinases
EC 2.7.7.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
2900Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM127094
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM127134
Pays : United States
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