Reconstitution of 30S ribosomal subunits in vitro using ribosome biogenesis factors

Daichi Tamaru, Kazuaki Amikura, Yoshihiro Shimizu, Knud H. Nierhaus, Takuya Ueda

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Reconstitution of ribosomes in vitro from individual ribosomal proteins provides a powerful tool for understanding the ribosome assembly process including the sequential incorporation of ribosomal proteins. However, conventional assembly methods require high-salt conditions for efficient ribosome assembly. In this study, we reconstituted 30S ribosomal subunits from individually purified ribosomal proteins in the presence of ribosome biogenesis factors. In this system, two GTPases (Era and YjeQ) facilitated assembly of a 30S subunit exhibiting poly(U)-directed polyphenylalanine synthesis and native protein synthesis under physiological conditions. This in vitro system permits a study of the assembly process and function of ribosome biogenesis factors, and it will facilitate the generation of ribosomes from DNA without using cells.

Original languageEnglish
Pages (from-to)1512-1519
Number of pages8
JournalRNA
Volume24
Issue number11
DOIs
Publication statusPublished - 2018 Nov

Keywords

  • Biogenesis factors
  • Cell-free translation system
  • In vitro reconstitution
  • Ribosome
  • Synthetic biology

ASJC Scopus subject areas

  • Molecular Biology

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