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TitleRibosome engineering reveals the importance of 5S rRNA autonomy for ribosome assembly.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 2900, Year 2020
Publish dateJun 9, 2020
AuthorsShijie Huang / Nikolay A Aleksashin / Anna B Loveland / Dorota Klepacki / Kaspar Reier / Amira Kefi / Teresa Szal / Jaanus Remme / Luc Jaeger / Nora Vázquez-Laslop / Andrei A Korostelev / Alexander S Mankin /
PubMed Abstract5S 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. ...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.
External linksNat Commun / PubMed:32518240 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 3.5 Å
Structure data

21856

6wnt

EMDB-21856, PDB-6wnt:
50S ribosomal subunit without free 5S rRNA and perturbed PTC
Method: EM (single particle) / Resolution: 3.1 Å

21857

6wnv

EMDB-21857, PDB-6wnv:
70S ribosome without free 5S rRNA and with a perturbed PTC
Method: EM (single particle) / Resolution: 3.5 Å

21858

6wnw

EMDB-21858, PDB-6wnw:
Active 70S ribosome without free 5S rRNA and bound with A- and P- tRNA
Method: EM (single particle) / Resolution: 3.2 Å

Source
  • escherichia coli (E. coli)
KeywordsRIBOSOME / 23S-cp5S rRNA / engineering

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