Engineered kinases as a tool for phosphorylation of selected targets in vivo

Lepeta, Katarzyna and Roubinet, Chantal and Bauer, Milena and Vigano, M. Alessandra and Aguilar, Gustavo and Kanca, Oguz and Ochoa-Espinosa, Amanda and Bieli, Dimitri and Cabernard, Clemens and Caussinus, Emmanuel and Affolter, Markus. (2022) Engineered kinases as a tool for phosphorylation of selected targets in vivo. Journal of Cell Biology, 221 (10). e202106179.

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Official URL: https://edoc.unibas.ch/89887/

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Reversible protein phosphorylation by kinases controls a plethora of processes essential for the proper development and homeostasis of multicellular organisms. One main obstacle in studying the role of a defined kinase-substrate interaction is that kinases form complex signaling networks and most often phosphorylate multiple substrates involved in various cellular processes. In recent years, several new approaches have been developed to control the activity of a given kinase. However, most of them fail to regulate a single protein target, likely hiding the effect of a unique kinase-substrate interaction by pleiotropic effects. To overcome this limitation, we have created protein binder-based engineered kinases that permit a direct, robust, and tissue-specific phosphorylation of fluorescent fusion proteins in vivo. We show the detailed characterization of two engineered kinases based on Rho-associated protein kinase (ROCK) and Src. Expression of synthetic kinases in the developing fly embryo resulted in phosphorylation of their respective GFP-fusion targets, providing for the first time a means to direct the phosphorylation to a chosen and tagged target in vivo. We presume that after careful optimization, the novel approach we describe here can be adapted to other kinases and targets in various eukaryotic genetic systems to regulate specific downstream effectors.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Growth & Development > Cell Biology (Affolter)
UniBasel Contributors:Affolter, Markus
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Rockefeller University Press
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:27 Sep 2022 08:33
Deposited On:27 Sep 2022 08:33

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