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A Yeast-Based Functional Assay to Study Plant N-Degron – N-Recognin Interactions

Kozlic, Aida; Winter, Nikola; Telser, Theresia; Reimann, Jakob; Rose, Katrin; Nehlin, Lilian; Berckhan, Sophie; Sharma, Gunjan; Dambire, Charlene; Boeckx, Tinne; Holdsworth, Michael J.; Bachmair, Andreas

A Yeast-Based Functional Assay to Study Plant N-Degron – N-Recognin Interactions Thumbnail


Authors

Aida Kozlic

Nikola Winter

Theresia Telser

Jakob Reimann

Katrin Rose

Lilian Nehlin

Sophie Berckhan

Gunjan Sharma

Charlene Dambire

Tinne Boeckx

Andreas Bachmair



Contributors

Gunjan Sharma
Researcher

Charlene Dambire
Researcher

Sophie Berckhan
Researcher

Abstract

The N-degron pathway is a branch of the ubiquitin-proteasome system where amino-terminal residues serve as degradation signals. In a synthetic biology approach, we expressed ubiquitin ligase PRT6 and ubiquitin conjugating enzyme 2 (AtUBC2) from Arabidopsis thaliana in a Saccharomyces cerevisiae strain with mutation in its endogenous N-degron pathway. The two enzymes re-constitute part of the plant N-degron pathway and were probed by monitoring the stability of co-expressed GFP-linked plant proteins starting with Arginine N-degrons. The novel assay allows for straightforward analysis, whereas in vitro interaction assays often do not allow detection of the weak binding of N-degron recognizing ubiquitin ligases to their substrates, and in planta testing is usually complex and time-consuming.

Citation

Kozlic, A., Winter, N., Telser, T., Reimann, J., Rose, K., Nehlin, L., Berckhan, S., Sharma, G., Dambire, C., Boeckx, T., Holdsworth, M. J., & Bachmair, A. (2022). A Yeast-Based Functional Assay to Study Plant N-Degron – N-Recognin Interactions. Frontiers in Plant Science, 12, Article 806129. https://doi.org/10.3389/fpls.2021.806129

Journal Article Type Article
Acceptance Date Dec 7, 2021
Online Publication Date Jan 7, 2022
Publication Date Jan 7, 2022
Deposit Date Jan 7, 2022
Publicly Available Date Jan 10, 2022
Journal Frontiers in Plant Science
Electronic ISSN 1664-462X
Publisher Frontiers Media
Peer Reviewed Peer Reviewed
Volume 12
Article Number 806129
DOI https://doi.org/10.3389/fpls.2021.806129
Keywords Plant Science
Public URL https://nottingham-repository.worktribe.com/output/7169669
Publisher URL https://www.frontiersin.org/articles/10.3389/fpls.2021.806129/full

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