Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance
Abstract Background The E. coli pET system is the most widely used protein over-expression system worldwide. It relies on the assumption that all cells produce target protein and it is generally believed that integral membrane protein (IMP) over-expression is more toxic than their soluble counterpar...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-01-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-020-01462-6 |
| _version_ | 1818960613735399424 |
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| author | Jack James Benjamin Yarnall Andy Koranteng Jane Gibson Tahmina Rahman Declan A. Doyle |
| author_facet | Jack James Benjamin Yarnall Andy Koranteng Jane Gibson Tahmina Rahman Declan A. Doyle |
| author_sort | Jack James |
| collection | DOAJ |
| description | Abstract Background The E. coli pET system is the most widely used protein over-expression system worldwide. It relies on the assumption that all cells produce target protein and it is generally believed that integral membrane protein (IMP) over-expression is more toxic than their soluble counterparts. Results Using GFP-tagged proteins, high level over-expression of either soluble or IMP targets results in > 99.9% cell loss with survival rate of only < 0.03%. Selective pressure generates three phenotypes: large green, large white and small colony variants. As a result, in overnight cultures, ~ 50% of the overall cell mass produces no protein. Genome sequencing of the phenotypes revealed genomic mutations that causes either the loss of T7 RNAP activity or its transcriptional downregulation. The over-expression process is bactericidal and is observed for both soluble and membrane proteins. Conclusions We demonstrate that it is the act of high-level over-expression of exogenous proteins in E. coli that sets in motion a chain of events leading to > 99.9% cell death. These results redefine our understanding of protein over-production and link it to the adaptive survival response seen in the development of antimicrobial resistance. |
| first_indexed | 2024-12-20T12:00:19Z |
| format | Article |
| id | doaj.art-64c69a0b96944ac1b61e1f6f988175f0 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-20T12:00:19Z |
| publishDate | 2021-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-64c69a0b96944ac1b61e1f6f988175f02022-12-21T19:41:33ZengBMCMicrobial Cell Factories1475-28592021-01-0120111110.1186/s12934-020-01462-6Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistanceJack James0Benjamin Yarnall1Andy Koranteng2Jane Gibson3Tahmina Rahman4Declan A. Doyle5School of Biological Sciences, University of SouthamptonSchool of Biological Sciences, University of SouthamptonSchool of Biological Sciences, University of SouthamptonCancer Sciences, Faculty of Medicine, University of SouthamptonQueen Alexandra Hospital, Portsmouth Hospital University NHS TrustSchool of Biological Sciences, University of SouthamptonAbstract Background The E. coli pET system is the most widely used protein over-expression system worldwide. It relies on the assumption that all cells produce target protein and it is generally believed that integral membrane protein (IMP) over-expression is more toxic than their soluble counterparts. Results Using GFP-tagged proteins, high level over-expression of either soluble or IMP targets results in > 99.9% cell loss with survival rate of only < 0.03%. Selective pressure generates three phenotypes: large green, large white and small colony variants. As a result, in overnight cultures, ~ 50% of the overall cell mass produces no protein. Genome sequencing of the phenotypes revealed genomic mutations that causes either the loss of T7 RNAP activity or its transcriptional downregulation. The over-expression process is bactericidal and is observed for both soluble and membrane proteins. Conclusions We demonstrate that it is the act of high-level over-expression of exogenous proteins in E. coli that sets in motion a chain of events leading to > 99.9% cell death. These results redefine our understanding of protein over-production and link it to the adaptive survival response seen in the development of antimicrobial resistance.https://doi.org/10.1186/s12934-020-01462-6Escherichia coliAntimicrobial resistanceBL21(DE3)Protein over-expression |
| spellingShingle | Jack James Benjamin Yarnall Andy Koranteng Jane Gibson Tahmina Rahman Declan A. Doyle Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance Microbial Cell Factories Escherichia coli Antimicrobial resistance BL21(DE3) Protein over-expression |
| title | Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance |
| title_full | Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance |
| title_fullStr | Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance |
| title_full_unstemmed | Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance |
| title_short | Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance |
| title_sort | protein over expression in escherichia coli triggers adaptation analogous to antimicrobial resistance |
| topic | Escherichia coli Antimicrobial resistance BL21(DE3) Protein over-expression |
| url | https://doi.org/10.1186/s12934-020-01462-6 |
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