Efficient protein depletion by genetically controlled deprotection of a dormant N-degron

• Verfasst von: Taxis, Christof [VerfasserIn]
• Verfasst von: Stier, Gunter [VerfasserIn]
• Verfasst von: Knop, Michael [VerfasserIn]
• Titel: Efficient protein depletion by genetically controlled deprotection of a dormant N-degron
• Titelzusatz: report
• Verf.angabe: Christof Taxis, Gunter Stier, Roberta Spadaccini, and Michael Knop
• E-Jahr: 2009
• Jahr: 2009 Apr 28
• Umfang: 7 S.
• Fussnoten: Gesehen am 14.08.2017
• Titel Quelle: Enthalten in: Molecular systems biology
• Ort Quelle: [London] : Nature Publishing Group UK, 2005
• Jahr Quelle: 2009
• Band/Heft Quelle: 5(2009) Article number 267, 7 Seiten
• ISSN Quelle: 1744-4292
• DOI: doi:10.1038/msb.2009.25
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1562393006

Abstract

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.