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

eLife | 2019

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.

Pubmed ID: 30950791 RIS Download

Associated grants

  • Agency: National Science Foundation, International
    Id: GRFP DGE-1122492
  • Agency: NCATS NIH HHS, United States
    Id: UL1 TR001863
  • Agency: National Science Foundation, International
    Id: EAGER #1548538
  • Agency: NIH HHS, United States
    Id: R35 GM128670
  • Agency: NIH HHS, United States
    Id: T32 GM007499
  • Agency: NIGMS NIH HHS, United States
    Id: T32 GM007499
  • Agency: NIGMS NIH HHS, United States
    Id: R35 GM128670

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This is a list of tools and resources that we have found mentioned in this publication.


UniProt (tool)

RRID:SCR_002380

Collection of data of protein sequence and functional information. Resource for protein sequence and annotation data. Consortium for preservation of the UniProt databases: UniProt Knowledgebase (UniProtKB), UniProt Reference Clusters (UniRef), and UniProt Archive (UniParc), UniProt Proteomes. Collaboration between European Bioinformatics Institute (EMBL-EBI), SIB Swiss Institute of Bioinformatics and Protein Information Resource. Swiss-Prot is a curated subset of UniProtKB.

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Promega (tool)

RRID:SCR_006724

An Antibody supplier

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Arabidopsis Biological Resource Center (tool)

RRID:SCR_008136

The mission of the Arabidopsis Biological Resource Center (ABRC) is to acquire, preserve and distribute seed and DNA resources that are useful to the Arabidopsis research community. More than 100,000 stocks are shipped annually to researchers in more than 60 countries, and modest fees for stocks are charged. The ABRC database functions and ordering system are incorporated into The Arabidopsis Information Resource (TAIR). Researchers can obtain information about Arabidopsis, perform stock searches, order stocks, and view current and past orders. Sponsors: ABRC is supported by the National Science Foundation under Grant No. 0542034.

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QIAGEN (tool)

RRID:SCR_008539

A commercial organization which provides assay technologies to isolate DNA, RNA, and proteins from any biological sample. Assay technologies are then used to make specific target biomolecules, such as the DNA of a specific virus, visible for subsequent analysis.

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MATLAB (software resource)

RRID:SCR_001622

Multi paradigm numerical computing environment and fourth generation programming language developed by MathWorks. Allows matrix manipulations, plotting of functions and data, implementation of algorithms, creation of user interfaces, and interfacing with programs written in other languages, including C, C++, Java, Fortran and Python. Used to explore and visualize ideas and collaborate across disciplines including signal and image processing, communications, control systems, and computational finance.

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ImageJ (software resource)

RRID:SCR_003070

Open source Java based image processing software program designed for scientific multidimensional images. ImageJ has been transformed to ImageJ2 application to improve data engine to be sufficient to analyze modern datasets.

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BRASS (software resource)

RRID:SCR_017091

Software tool for analysis of one or more related BAM files of paired end sequencing to determine potential rearrangement breakpoints. Identifies breaks and attempts to assemble rearrangements.

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MATLAB (software resource)

RRID:SCR_001622

Multi paradigm numerical computing environment and fourth generation programming language developed by MathWorks. Allows matrix manipulations, plotting of functions and data, implementation of algorithms, creation of user interfaces, and interfacing with programs written in other languages, including C, C++, Java, Fortran and Python. Used to explore and visualize ideas and collaborate across disciplines including signal and image processing, communications, control systems, and computational finance.

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BRASS (software resource)

RRID:SCR_017091

Software tool for analysis of one or more related BAM files of paired end sequencing to determine potential rearrangement breakpoints. Identifies breaks and attempts to assemble rearrangements.

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ImageJ (software resource)

RRID:SCR_003070

Open source Java based image processing software program designed for scientific multidimensional images. ImageJ has been transformed to ImageJ2 application to improve data engine to be sufficient to analyze modern datasets.

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