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doi:10.22028/D291-43706
Titel: | Metabolite-responsive control of transcription by phase separation-based synthetic organelles |
VerfasserIn: | Jerez-Longres, Carolina Weber, Wilfried |
Sprache: | Englisch |
Verlag/Plattform: | bioRxiv |
Erscheinungsjahr: | 2024 |
Freie Schlagwörter: | Coacervate intrinsically disordered region in vitro transcription liquid-liquid phase separation repressor protein |
DDC-Sachgruppe: | 620 Ingenieurwissenschaften und Maschinenbau |
Dokumenttyp: | Sonstiges |
Abstract: | Living natural materials have remarkable sensing abilities that translate external cues into functional changes of the material. The reconstruction of such sensing materials in bottom-up synthetic biology provides the opportunity to develop synthetic materials with life-like sensing and adaptation ability. Key to such functions are material modules that translate specific input signals into a biomolecular response. Here, we engineer a synthetic organelle based on liquid-liquid phase separation that translates a metabolic signal into the regulation of gene transcription. To this aim, we engineer the pyruvate-dependent repressor PdhR to undergo liquid-liquid phase separation in vitro by fusion to intrinsically disordered regions. We demonstrate that the resulting coacervates bind DNA harbouring PdhR-responsive operator sites in a pyruvate dose-dependent and reversible manner. We observed that the activity of transcription units on the DNA was strongly attenuated following recruitment to the coacervates. However, the addition of pyruvate resulted in a reversible and dose-dependent reconstitution of transcriptional activity. The coacervate-based synthetic organelles linking metabolic cues to transcriptional signals represent a materials approach to confer stimulus-responsiveness to minimal bottom-up synthetic biological systems and open opportunities in materials for sensor applications. |
DOI der Erstveröffentlichung: | 10.1101/2024.09.17.613456 |
URL der Erstveröffentlichung: | https://www.biorxiv.org/content/10.1101/2024.09.17.613456v1 |
Link zu diesem Datensatz: | urn:nbn:de:bsz:291--ds-437066 hdl:20.500.11880/39148 http://dx.doi.org/10.22028/D291-43706 |
Datum des Eintrags: | 10-Dez-2024 |
Bemerkung/Hinweis: | Preprint |
Fakultät: | NT - Naturwissenschaftlich- Technische Fakultät |
Fachrichtung: | NT - Materialwissenschaft und Werkstofftechnik |
Professur: | NT - Prof. Dr. Wilfried Weber |
Sammlung: | SciDok - Der Wissenschaftsserver der Universität des Saarlandes |
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