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Elimination of Ribosome Inactivating Factors Improves the Efficiency of Bacillus subtilis and Saccharomyces cerevisiae Cell-Free Translation Systems
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Institute of Technology, University of Tartu, Tartu, Estonia.
Umeå University, Faculty of Science and Technology, Department of Molecular Biology (Faculty of Science and Technology).
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine). Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS).
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2018 (English)In: Frontiers in Microbiology, ISSN 1664-302X, E-ISSN 1664-302X, Vol. 9, article id 3041Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
Frontiers Media S.A., 2018. Vol. 9, article id 3041
Keywords [en]
HPF, Stm1, Bacillus subtilis, Saccharomyces cerevisiae, cell-tree translation system
National Category
Microbiology
Identifiers
URN: urn:nbn:se:umu:diva-155100DOI: 10.3389/fmicb.2018.03041ISI: 000453653000001OAI: oai:DiVA.org:umu-155100DiVA, id: diva2:1277273
Funder
Swedish Research Council, 2013-4680Swedish Research Council, 2017-04663Ragnar Söderbergs stiftelseMagnus Bergvall Foundation, 2017-02098Åke Wiberg Foundation, M14-0207Available from: 2019-01-10 Created: 2019-01-10 Last updated: 2019-01-10Bibliographically approved

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Johansson, Marcus J. O.Takada, HirakuHauryliuk, VasiliMurina, Victoriia

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Johansson, Marcus J. O.Takada, HirakuHauryliuk, VasiliMurina, Victoriia
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Molecular Infection Medicine Sweden (MIMS)Department of Molecular Biology (Faculty of Science and Technology)Department of Molecular Biology (Faculty of Medicine)
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Frontiers in Microbiology
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