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A bacterial type III secretion-based protein delivery tool for broad applications in cell biology

Ittig, Simon Josef and Schmutz, Christoph and Kasper, Christoph Alexander and Amstutz, Marlise and Schmidt, Alexander and Sauteur, Loïc and Vigano, M. Alessandra and Low, Shyan Huey and Affolter, Markus and Cornelis, Guy R. and Nigg, Erich A. and Arrieumerlou, Cécile. (2015) A bacterial type III secretion-based protein delivery tool for broad applications in cell biology. The Journal of cell biology, 211 (4). pp. 913-931.

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Official URL: http://edoc.unibas.ch/39906/

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Abstract

Methods enabling the delivery of proteins into eukaryotic cells are essential to address protein functions. Here we propose broad applications to cell biology for a protein delivery tool based on bacterial type III secretion (T3S). We show that bacterial, viral, and human proteins, fused to the N-terminal fragment of the Yersinia enterocolitica T3S substrate YopE, are effectively delivered into target cells in a fast and controllable manner via the injectisome of extracellular bacteria. This method enables functional interaction studies by the simultaneous injection of multiple proteins and allows the targeting of proteins to different subcellular locations by use of nanobody-fusion proteins. After delivery, proteins can be freed from the YopE fragment by a T3S-translocated viral protease or fusion to ubiquitin and cleavage by endogenous ubiquitin proteases. Finally, we show that this delivery tool is suitable to inject proteins in living animals and combine it with phosphoproteomics to characterize the systems-level impact of proapoptotic human truncated BID on the cellular network.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Growth & Development > Cell Biology (Affolter)
05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Cell Biology (Nigg)
UniBasel Contributors:Affolter, Markus and Nigg, Erich A. and Ittig, Simon Josef and Schmutz, Christoph and Kasper, Christoph and Amstutz, Marlise and Schmidt, Alexander and Sauteur, LoÏc and Vigano, Maria Alessandra and Low, Shyan Huey and Arrieumerlou, Cécile
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Rockefeller University Press
ISSN:0021-9525
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:30 Jun 2016 11:00
Deposited On:30 May 2016 06:34

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