Reconstitution of 30S ribosomal subunits in vitro using ribosome biogenesis factors
- 1 The University of Tokyo;
- 2 RIKEN Quantitative Biology Center;
- 3 Charité-Universitätsmedizin Berlin
- ↵* Corresponding author; email: amikura{at}k.u-tokyo.ac.jp
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 30S subunit exhibiting poly(U)-directed polyphenylalanine synthesis and native protein, under physiological condition. This in vitro system permits study of the assembly process and function of ribosome biogenesis factors, and it will facilitate the generation of ribosomes from DNA without using cells.
Keywords
- Received March 2, 2018.
- Accepted July 25, 2018.
- Published by Cold Spring Harbor Laboratory Press for the RNA Society
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