AUTHOR=Brodiazhenko Tetiana , Johansson Marcus J. O. , Takada Hiraku , Nissan Tracy , Hauryliuk Vasili , Murina Victoriia 

TITLE=Elimination of Ribosome Inactivating Factors Improves the Efficiency of Bacillus subtilis and Saccharomyces cerevisiae Cell-Free Translation Systems

JOURNAL=Frontiers in Microbiology

VOLUME=9

YEAR=2018

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2018.03041

DOI=10.3389/fmicb.2018.03041

ISSN=1664-302X

ABSTRACT=<p>Cell-free translation systems based on cellular lysates optimized for <italic>in vitro</italic> 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 <italic>Bacillus subtilis</italic> and Stm1 in <italic>Saccharomyces cerevisiae</italic> – robustly improve the activities of bacterial and yeast translation systems. Importantly, the elimination of <italic>B. subtilis</italic> 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.</p>