In vitro reconstitution of functional small ribosomal subunit assembly for comprehensive analysis of ribosomal elements in E. coli

Masaru Shimojo, Kazuaki Amikura, Keiko Masuda, Takashi Kanamori, Takuya Ueda, Yoshihiro Shimizu*

*この研究の対応する著者

研究成果: Article査読

11 被引用数 (Scopus)

抄録

In vitro reconstitution is a powerful tool for investigating ribosome functions and biogenesis, as well as discovering new ribosomal features. In this study, we integrated all of the processes required for Escherichia coli small ribosomal subunit assembly. In our method, termed fully Recombinant-based integrated Synthesis, Assembly, and Translation (R-iSAT), assembly and evaluation of the small ribosomal subunits are coupled with ribosomal RNA (rRNA) synthesis in a reconstituted cell-free protein synthesis system. By changing the components of R-iSAT, including recombinant ribosomal protein composition, we coupled ribosomal assembly with ribosomal protein synthesis, enabling functional synthesis of ribosomal proteins and subsequent subunit assembly. In addition, we assembled and evaluated subunits with mutations in both rRNA and ribosomal proteins. The study demonstrated that our scheme provides new ways to comprehensively analyze any elements of the small ribosomal subunit, with the goal of improving our understanding of ribosomal biogenesis, function, and engineering.

本文言語English
論文番号142
ジャーナルCommunications Biology
3
1
DOI
出版ステータスPublished - 2020 12月 1

ASJC Scopus subject areas

  • 医学(その他)
  • 生化学、遺伝学、分子生物学(全般)
  • 農業および生物科学(全般)

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