System‐wide optimization of an orthogonal translation system with enhanced biological tolerance
Abstract Over the past two decades, synthetic biological systems have revolutionized the study of cellular physiology. The ability to site‐specifically incorporate biologically relevant non‐standard amino acids using orthogonal translation systems (OTSs) has proven particularly useful, providing unp...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Springer Nature
2023-07-01
|
| Series: | Molecular Systems Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.15252/msb.202110591 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1846171528523153408 |
|---|---|
| author | Kyle Mohler Jack M Moen Svetlana Rogulina Jesse Rinehart |
| author_facet | Kyle Mohler Jack M Moen Svetlana Rogulina Jesse Rinehart |
| author_sort | Kyle Mohler |
| collection | DOAJ |
| description | Abstract Over the past two decades, synthetic biological systems have revolutionized the study of cellular physiology. The ability to site‐specifically incorporate biologically relevant non‐standard amino acids using orthogonal translation systems (OTSs) has proven particularly useful, providing unparalleled access to cellular mechanisms modulated by post‐translational modifications, such as protein phosphorylation. However, despite significant advances in OTS design and function, the systems‐level biology of OTS development and utilization remains underexplored. In this study, we employ a phosphoserine OTS (pSerOTS) as a model to systematically investigate global interactions between OTS components and the cellular environment, aiming to improve OTS performance. Based on this analysis, we design OTS variants to enhance orthogonality by minimizing host process interactions and reducing stress response activation. Our findings advance understanding of system‐wide OTS:host interactions, enabling informed design practices that circumvent deleterious interactions with host physiology while improving OTS performance and stability. Furthermore, our study emphasizes the importance of establishing a pipeline for systematically profiling OTS:host interactions to enhance orthogonality and mitigate mechanisms underlying OTS‐mediated host toxicity. |
| format | Article |
| id | doaj-art-85d65118eae1479585f4602431d54ea4 |
| institution | Kabale University |
| issn | 1744-4292 |
| language | English |
| publishDate | 2023-07-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-85d65118eae1479585f4602431d54ea42024-11-10T12:48:28ZengSpringer NatureMolecular Systems Biology1744-42922023-07-0119811810.15252/msb.202110591System‐wide optimization of an orthogonal translation system with enhanced biological toleranceKyle Mohler0Jack M Moen1Svetlana Rogulina2Jesse Rinehart3Department of Cellular & Molecular Physiology, Yale School of MedicineQuantitative Biosciences Institute (QBI), University of California, San FranciscoDepartment of Cellular & Molecular Physiology, Yale School of MedicineDepartment of Cellular & Molecular Physiology, Yale School of MedicineAbstract Over the past two decades, synthetic biological systems have revolutionized the study of cellular physiology. The ability to site‐specifically incorporate biologically relevant non‐standard amino acids using orthogonal translation systems (OTSs) has proven particularly useful, providing unparalleled access to cellular mechanisms modulated by post‐translational modifications, such as protein phosphorylation. However, despite significant advances in OTS design and function, the systems‐level biology of OTS development and utilization remains underexplored. In this study, we employ a phosphoserine OTS (pSerOTS) as a model to systematically investigate global interactions between OTS components and the cellular environment, aiming to improve OTS performance. Based on this analysis, we design OTS variants to enhance orthogonality by minimizing host process interactions and reducing stress response activation. Our findings advance understanding of system‐wide OTS:host interactions, enabling informed design practices that circumvent deleterious interactions with host physiology while improving OTS performance and stability. Furthermore, our study emphasizes the importance of establishing a pipeline for systematically profiling OTS:host interactions to enhance orthogonality and mitigate mechanisms underlying OTS‐mediated host toxicity.https://doi.org/10.15252/msb.202110591bacterial physiologyorthogonal translation systemphosphoserinestress tolerancesynthetic biology |
| spellingShingle | Kyle Mohler Jack M Moen Svetlana Rogulina Jesse Rinehart System‐wide optimization of an orthogonal translation system with enhanced biological tolerance Molecular Systems Biology bacterial physiology orthogonal translation system phosphoserine stress tolerance synthetic biology |
| title | System‐wide optimization of an orthogonal translation system with enhanced biological tolerance |
| title_full | System‐wide optimization of an orthogonal translation system with enhanced biological tolerance |
| title_fullStr | System‐wide optimization of an orthogonal translation system with enhanced biological tolerance |
| title_full_unstemmed | System‐wide optimization of an orthogonal translation system with enhanced biological tolerance |
| title_short | System‐wide optimization of an orthogonal translation system with enhanced biological tolerance |
| title_sort | system wide optimization of an orthogonal translation system with enhanced biological tolerance |
| topic | bacterial physiology orthogonal translation system phosphoserine stress tolerance synthetic biology |
| url | https://doi.org/10.15252/msb.202110591 |
| work_keys_str_mv | AT kylemohler systemwideoptimizationofanorthogonaltranslationsystemwithenhancedbiologicaltolerance AT jackmmoen systemwideoptimizationofanorthogonaltranslationsystemwithenhancedbiologicaltolerance AT svetlanarogulina systemwideoptimizationofanorthogonaltranslationsystemwithenhancedbiologicaltolerance AT jesserinehart systemwideoptimizationofanorthogonaltranslationsystemwithenhancedbiologicaltolerance |