Efficient protein depletion by genetically controlled deprotection of a dormant N-degron: report
Methods that allow for the manipulation of genes or their products have been highly fruitful for biomedical research. Here, we describe a method that allows the control of protein abundance by a genetically encoded regulatory system. We developed a dormant N-degron that can be attached to the N-term...
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| Hauptverfasser: | , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
2009 Apr 28
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| In: |
Molecular systems biology
Year: 2009, Jahrgang: 5 |
| ISSN: | 1744-4292 |
| DOI: | 10.1038/msb.2009.25 |
| Online-Zugang: | Verlag, kostenfrei, Volltext: http://dx.doi.org/10.1038/msb.2009.25 Verlag, kostenfrei, Volltext: https://www.embopress.org/doi/full/10.1038/msb.2009.25 
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| Verfasserangaben: | Christof Taxis, Gunter Stier, Roberta Spadaccini, and Michael Knop |
| Zusammenfassung: | Methods that allow for the manipulation of genes or their products have been highly fruitful for biomedical research. Here, we describe a method that allows the control of protein abundance by a genetically encoded regulatory system. We developed a dormant N-degron that can be attached to the N-terminus of a protein of interest. Upon expression of a site-specific protease, the dormant N-degron becomes deprotected. The N-degron then targets itself and the attached protein for rapid proteasomal degradation through the N-end rule pathway. We use an optimized tobacco etch virus (TEV) protease variant combined with selective target binding to achieve complete and rapid deprotection of the N-degron-tagged proteins. This method, termed TEV protease induced protein inactivation (TIPI) of TIPI-degron (TDeg) modified target proteins is fast, reversible, and applicable to a broad range of proteins. TIPI of yeast proteins essential for vegetative growth causes phenotypes that are close to deletion mutants. The features of the TIPI system make it a versatile tool to study protein function in eukaryotes and to create new modules for synthetic or systems biology. |
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| Beschreibung: | Gesehen am 14.08.2017 |
| Beschreibung: | Online Resource |
| ISSN: | 1744-4292 |
| DOI: | 10.1038/msb.2009.25 |