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Updated: Apr 4, 2026

Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta
Published on: December 5, 2019
The RING E3 Ligase KEEP ON GOING Modulates JASMONATE ZIM-DOMAIN12 Stability
Laurens Pauwels1, Andrés Ritter1, Jonas Goossens1
1Department of Plant Systems Biology, Flanders Institute for Biotechnology, B-9052 Ghent, Belgium (L.P., A.R., J.Go., A.N.D., J.Ge., R.V.B., R.D.C., J.V.L., G.D.J., A.G.);Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (L.P., A.R., J.Go., A.N.D., J.Ge., R.V.B., R.D.C., J.V.L., G.D.J., A.G.);Department of Molecular and Cellular Biology, University of California, Davis, California 95616 (L.P., J.C.);Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4R2 (H.L., S.S.);Department of Biology, Indiana University, Bloomington, Indiana 47405 (Y.Gu, R.W.I.);Plant Molecular Genetics Department, National Centre for Biotechnology, Consejo Superior de Investigaciones Científicas, Campus University Autónoma, 28049 Madrid, Spain (M.B., R.S.);Department of Medical Protein Research, Flanders Institute for Biotechnology, 9000 Ghent, Belgium (K.G.); andDepartment of Biochemistry, Ghent University, 9000 Ghent, Belgium (K.G.).
Jasmonate signaling involves JAZ proteins, degraded via COI1. This study reveals JAZ12 interacts with KEG, a ligase regulating JAZ12 stability and plant hormone responses.
Area of Science:
- Plant Molecular Biology
- Plant Hormone Signaling
Background:
- Jasmonate (JA) signaling is crucial for plant development and defense.
- JASMONATE ZIM-DOMAIN (JAZ) proteins mediate JA signaling by repressing transcription factors.
- The F-box protein CORONATINE INSENSITIVE1 (COI1) targets JAZ proteins for degradation.
Purpose of the Study:
- To investigate the function and regulation of JAZ12, a highly expressed JAZ protein in Arabidopsis.
- To explore the interaction between JAZ12 and the E3 RING ligase KEEP ON GOING (KEG).
Main Methods:
- Yeast two-hybrid assays and tandem affinity purification to identify protein interactions.
- In vivo ubiquitination assays and Western blotting to assess protein degradation.
- Artificial microRNA to silence KEG in Arabidopsis to circumvent mutant lethality.
Main Results:
- JAZ12 interacts with MYC2, MYC3, and MYC4, repressing MYC2 activity.
- JAZ12 interacts with SCF(COI1) components, leading to JA-induced degradation.
- JAZ12 directly interacts with KEG, influencing its stability.
- KEG knockdown reduces JAZ12 protein levels, while KEG overexpression inhibits COI1-mediated JAZ12 degradation.
Conclusions:
- JAZ12 plays a role in JA signaling by interacting with MYC transcription factors.
- KEG positively regulates JAZ12 stability and is involved in plant hormone signaling pathways.
- The interaction between JAZ12 and KEG adds complexity to the regulation of JA signaling.
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