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Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function

The circadian clock relies on regulated degradation of clock proteins to maintain rhythmicity. Despite this, we know few components that mediate protein degradation. This is due to high levels of functional redundancy within plant E3 ubiquitin ligase families. In order to overcome this issue and dis...

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Autores principales: Feke, Ann, Liu, Wei, Hong, Jing, Li, Man-Wah, Lee, Chin-Mei, Zhou, Elton K, Gendron, Joshua M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6483598/
https://www.ncbi.nlm.nih.gov/pubmed/30950791
http://dx.doi.org/10.7554/eLife.44558
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author Feke, Ann
Liu, Wei
Hong, Jing
Li, Man-Wah
Lee, Chin-Mei
Zhou, Elton K
Gendron, Joshua M
author_facet Feke, Ann
Liu, Wei
Hong, Jing
Li, Man-Wah
Lee, Chin-Mei
Zhou, Elton K
Gendron, Joshua M
author_sort Feke, Ann
collection PubMed
description The circadian clock relies on regulated degradation of clock proteins to maintain rhythmicity. Despite this, we know few components that mediate protein degradation. This is due to high levels of functional redundancy within plant E3 ubiquitin ligase families. In order to overcome this issue and discover E3 ubiquitin ligases that control circadian function, we generated a library of transgenic Arabidopsis plants expressing dominant-negative ‘decoy’ E3 ubiquitin ligases. We determined their effects on the circadian clock and identified dozens of new potential regulators of circadian function. To demonstrate the potency of the decoy screening methodology to overcome redundancy and identify bona fide clock regulators, we performed follow-up studies on MAC3A (PUB59) and MAC3B (PUB60). We show that they redundantly control circadian period by regulating splicing. This work demonstrates the viability of ubiquitin ligase decoys as a screening platform to overcome genetic challenges and discover E3 ubiquitin ligases that regulate plant development.
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spelling pubmed-64835982019-04-29 Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function Feke, Ann Liu, Wei Hong, Jing Li, Man-Wah Lee, Chin-Mei Zhou, Elton K Gendron, Joshua M eLife Plant Biology The circadian clock relies on regulated degradation of clock proteins to maintain rhythmicity. Despite this, we know few components that mediate protein degradation. This is due to high levels of functional redundancy within plant E3 ubiquitin ligase families. In order to overcome this issue and discover E3 ubiquitin ligases that control circadian function, we generated a library of transgenic Arabidopsis plants expressing dominant-negative ‘decoy’ E3 ubiquitin ligases. We determined their effects on the circadian clock and identified dozens of new potential regulators of circadian function. To demonstrate the potency of the decoy screening methodology to overcome redundancy and identify bona fide clock regulators, we performed follow-up studies on MAC3A (PUB59) and MAC3B (PUB60). We show that they redundantly control circadian period by regulating splicing. This work demonstrates the viability of ubiquitin ligase decoys as a screening platform to overcome genetic challenges and discover E3 ubiquitin ligases that regulate plant development. eLife Sciences Publications, Ltd 2019-04-05 /pmc/articles/PMC6483598/ /pubmed/30950791 http://dx.doi.org/10.7554/eLife.44558 Text en © 2019, Feke et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Plant Biology
Feke, Ann
Liu, Wei
Hong, Jing
Li, Man-Wah
Lee, Chin-Mei
Zhou, Elton K
Gendron, Joshua M
Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
title Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
title_full Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
title_fullStr Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
title_full_unstemmed Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
title_short Decoys provide a scalable platform for the identification of plant E3 ubiquitin ligases that regulate circadian function
title_sort decoys provide a scalable platform for the identification of plant e3 ubiquitin ligases that regulate circadian function
topic Plant Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6483598/
https://www.ncbi.nlm.nih.gov/pubmed/30950791
http://dx.doi.org/10.7554/eLife.44558
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