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Front. Microbiol. | doi: 10.3389/fmicb.2018.03041

Elimination of ribosome inactivating factors improves the efficiency of Bacillus subtilis and Saccharomyces cerevisiae cell-free translational systems

Tetiana Brodiazhenko1, Marcus J. Johansson2,  Hiraku Takada2,  Tracy Nissan3,  Vasili Hauryliuk2* and  Victoriia Murina2*
  • 1University of Tartu, Estonia
  • 2Umeå University, Sweden
  • 3University of Sussex, United Kingdom

Cell-free translation systems based on cellular lysates optimized for in vitro protein synthesis have multiple applications both in basic and applied science, ranging from studies of translational regulation to cell-free production of proteins and ribosome-nascent chain complexes. In order to achieve both high activity and reproducibility in a translation system, it is essential that the ribosomes in the cellular lysate are enzymatically active. Here we demonstrate that genomic disruption of genes encoding ribosome inactivating factors – HPF in Bacillus subtilis and Stm1 in Saccharomyces cerevisiae – robustly improve the activities of bacterial and yeast translation systems. Importantly, the elimination of B. subtilis HPF results in a complete loss of 100S ribosomes, which otherwise interfere with disome-based approaches for preparation of stalled ribosomal complexes for cryo-electron microscopy studies.

Keywords: HPF, Stm1, Bacillus subtilis (B. subtilis), Saccharomyces cerevisiae (Baker's yeast), Cell-free translational system

Received: 09 Oct 2018; Accepted: 26 Nov 2018.

Edited by:

Reynald GILLET, University of Rennes 1, France

Reviewed by:

Hyouta Himeno, Hirosaki University, Japan
Yoshihiro Shimizu, RIKEN, Japan  

Copyright: © 2018 Brodiazhenko, Johansson, Takada, Nissan, Hauryliuk and Murina. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Prof. Vasili Hauryliuk, Umeå University, Umeå, Sweden, vasili.hauryliuk@umu.se
Dr. Victoriia Murina, Umeå University, Umeå, Sweden, victoriia.murina@umu.se