MazF activation causes ACA sequence-independent and selective alterations in RNA levels in Escherichia coli

Akiyama, Kentaro; Fujisawa, Kazuki; Kondo, Hiro; Netsu, Yuya; Nishikawa, Koji; Takata, Yoshio; Nakamura, Yuya; Kino, Yuta; Ayukawa, Shotaro; Yamamura, Masatoshi; Hayashi, Nobuhiro; Tagawa, Yoh‐ichi; Nakashima, Naotoshi

doi:10.1007/s00203-019-01726-9

Cited by 2 publications

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References 47 publications

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“…MazF acts as an endoribonuclease that cleaves single-stranded RNA regions at ACA trinucleotide sites in Escherichia coli [3,4]. As artificially produced MazF degrades the rRNA precursors, as well as mRNAs [4][5][6][7][8], overall translation is reduced, which leads to decline in bacterial growth [9]. Even though cells grow slowly during mazF overexpression, cellular processes such as transcription and translation are not halted during this growth reduction [10,11].…”

Section: Introductionmentioning

confidence: 99%

Quantifying heterologous gene expression during ectopic MazF production in Escherichia coli

et al. 2022

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Objective MazF is a sequence-specific endoribonuclease-toxin of the MazEF toxin–antitoxin system. MazF cleaves single-stranded ribonucleic acid (RNA) regions at adenine–cytosine–adenine (ACA) sequences in the bacterium Escherichia coli. The MazEF system has been used in various biotechnology and synthetic biology applications. In this study, we infer how ectopic mazF overexpression affects production of heterologous proteins. To this end, we quantified the levels of fluorescent proteins expressed in E. coli from reporters translated from the ACA-containing or ACA-less messenger RNAs (mRNAs). Additionally, we addressed the impact of the 5′-untranslated region of these reporter mRNAs under the same conditions by comparing expression from mRNAs that comprise (canonical mRNA) or lack this region (leaderless mRNA). Results Flow cytometry analysis indicates that during mazF overexpression, fluorescent proteins are translated from the canonical as well as leaderless mRNAs. Our analysis further indicates that longer mazF overexpression generally increases the concentration of fluorescent proteins translated from ACA-less mRNAs, however it also substantially increases bacterial population heterogeneity. Finally, our results suggest that the strength and duration of mazF overexpression should be optimized for each experimental setup, to maximize the heterologous protein production and minimize the amount of phenotypic heterogeneity in bacterial populations, which is unfavorable in biotechnological processes.

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Section: Introductionmentioning

confidence: 99%

Quantifying heterologous gene expression during ectopic MazF production in Escherichia coli

et al. 2022

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“…We hypothesize that the decrease in the expression of mazEF in the pMUT strains would affect the ability of MazEF to carry out its biological function. However, the biological function of MazEF remains undefined ( 20 , 42 – 45 ). Recently, a study globally identified proteins, including the DNA-binding protein from starved cells (Dps), that were upregulated in a wild-type strain compared to a mazEF deletion strain after DNA damage ( 42 ).…”

Section: Resultsmentioning

confidence: 99%

An Antisense RNA Fine-Tunes Gene Expression of the Type II MazEF Toxin-Antitoxin System

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Martin

Bono

et al. 2022

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MazF activation causes ACA sequence-independent and selective alterations in RNA levels in Escherichia coli

Cited by 2 publications

References 47 publications

Quantifying heterologous gene expression during ectopic MazF production in Escherichia coli

Quantifying heterologous gene expression during ectopic MazF production in Escherichia coli

An Antisense RNA Fine-Tunes Gene Expression of the Type II MazEF Toxin-Antitoxin System

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