Data_Sheet_4_A Yeast-Based Functional Assay to Study Plant N-Degron – N-Recognin Interactions.pdf
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.
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References
- https://doi.org//10.1126/science.3018930
- https://doi.org//10.1146/annurev-arplant-050718-095937
- https://doi.org//10.1016/j.febslet.2007.06.005
- https://doi.org//10.1038/nature10534
- https://doi.org//10.1038/s41467-018-07875-7
- https://doi.org//10.1016/0378-1119%2888%2990185-0
- https://doi.org//10.1104/pp.19.00251
- https://doi.org//10.1016/j.molcel.2010.12.022
- https://doi.org//10.1111/tpj.13603
- https://doi.org//10.1111/jipb.12882
- https://doi.org//10.1111/nph.14900
- https://doi.org//10.1016/s0092-8674%2803%2900040-0
- https://doi.org//10.1016/0378-1119%2883%2990126-9
- https://doi.org//10.1146/annurev-arplant-050718-100211
- https://doi.org//10.1111/nph.14497
- https://doi.org//10.1016/j.jmb.2019.05.023
- https://doi.org//10.1186/s12870-015-0660-9
- https://doi.org//10.1104/pp.103.029272
- https://doi.org//10.1104/pp.104.052423
- https://doi.org//10.1186/1471-2229-9-39
- https://doi.org//10.1002/yea.320101310
- https://doi.org//10.1038/s41586-019-1203-6
- https://doi.org//10.1038/ncomms14690
- https://doi.org//10.1104/pp.19.00051
- https://doi.org//10.1111/tpj.12127