Elimination of Ribosome Inactivating Factors Improves the Efficiency of Bacillus subtilis and Saccharomyces cerevisiae Cell-Free Translation Systems
Brodiazhenko, Tetiana ; Johansson, Marcus J O LU ; Takada, Hiraku ; Nissan, Tracy ; Hauryliuk, Vasili LU
and Murina, Victoriia
(2018)
In Frontiers in Microbiology
9.
- Abstract
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... (More)
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.
(Less)
- author
- Brodiazhenko, Tetiana
; Johansson, Marcus J O
LU
; Takada, Hiraku
; Nissan, Tracy
; Hauryliuk, Vasili
LU
and Murina, Victoriia
- publishing date
- 2018
- type
- Contribution to journal
- publication status
- published
- keywords
- Bacillus subtilis, cell-free translation system, HPF, Saccharomyces cerevisiae, Stm1
- in
- Frontiers in Microbiology
- volume
- 9
- article number
- 3041
- publisher
- Frontiers Media S. A.
- external identifiers
-
- scopus:85082559395
- pmid:30619132
- ISSN
- 1664-302X
- DOI
- 10.3389/fmicb.2018.03041
- language
- English
- LU publication?
- no
- id
- c30da890-c799-4d3e-9c6f-f1793dc1e8ee
- date added to LUP
- 2021-11-10 12:51:43
- date last changed
- 2024-04-20 15:54:31
@article{c30da890-c799-4d3e-9c6f-f1793dc1e8ee,
abstract = {{<p>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.</p>}},
author = {{Brodiazhenko, Tetiana and Johansson, Marcus J O and Takada, Hiraku and Nissan, Tracy and Hauryliuk, Vasili and Murina, Victoriia}},
issn = {{1664-302X}},
keywords = {{Bacillus subtilis; cell-free translation system; HPF; Saccharomyces cerevisiae; Stm1}},
language = {{eng}},
publisher = {{Frontiers Media S. A.}},
series = {{Frontiers in Microbiology}},
title = {{Elimination of Ribosome Inactivating Factors Improves the Efficiency of Bacillus subtilis and Saccharomyces cerevisiae Cell-Free Translation Systems}},
url = {{http://dx.doi.org/10.3389/fmicb.2018.03041}},
doi = {{10.3389/fmicb.2018.03041}},
volume = {{9}},
year = {{2018}},
}