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

Feke, Ann; Liu, Wei; Hong, Jing Han; Li, Man-Wah; Lee, Chin-Mei; Zhou, Elton; Gendron, Joshua M.

doi:10.7554/elife.44558

Cited by 30 publications

(44 citation statements)

References 120 publications

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“…We have noted previously that complex formation between E3 ubiquitin ligases and their homologs may be a common feature of this class of proteins, and the ability of ZTL, FKF1, and LKP2 to heterodimerize has been previously reported [13,14,31,36,37]. We demonstrate that the homodimerization takes place between the LOV and Kelch repeat domains, suggesting that the interactions between ZTL and FKF1/LKP2 may also occur in this manner.…”

Section: Ztl Kelch Repeat Interaction Profilessupporting

confidence: 63%

“…We included CCA1p::Luciferase plants that express FKF1, LKP2, and ZTL decoys (LOV-Kelch fusion proteins which lack the F-box domain), which we have analyzed previously [13], and wild type CCA1p::Luciferase parental plants, as controls. In order to compare results from experiments performed separately, we use the difference between the period or flowering time of the individual T1 transgenic and the average period or flowering time of the concurrent wild type control plants for our statistical analyses [36,37]. The data generated in these experiments is displayed in Figure 1 and Tables 1 and 2.…”

Section: Expression Of the Lov Domain Of Lov/f-box/kelch Proteins Dismentioning

confidence: 99%

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Functional domain studies uncover novel roles for the ZTL Kelch repeat domain in clock function

Feke

Gendron

2020

Preprint

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The small LOV/F-box/Kelch family of E3 ubiquitin ligases plays an essential role in the regulation of plant circadian clocks and flowering time by sensing dusk. The family consists of three members, ZEITLUPE (ZTL), LOV KELCH PROTEIN 2 (LKP2), and FLAVIN-BINDING KELCH REPEAT F-BOX PROTEIN 1 (FKF1), which share a unique protein domain architecture allowing them to act as photoreceptors that transduce light signals via altering stability of target proteins. Despite intensive study of this protein family we still lack important knowledge about the biochemical and functional roles of the protein domains that comprise these unique photoreceptors. Here, we perform comparative analyses of transgenic lines constitutively expressing the photoreceptor LOV domain or the Kelch repeat protein-protein interaction domains of ZTL, FKF1, and LKP2. Expression of each domain alone is sufficient to disrupt circadian rhythms and flowering time, but each domain differs in the magnitude of effect. Immunoprecipitation followed by mass spectrometry with the ZTL Kelch repeat domain identified a suite of potential interacting partners. Furthermore, the ZTL Kelch repeat domain mediates interaction with the LOV domain of ZTL and the ZTL homologs LKP2 and FKF1. This suggests that the Kelch repeat domain of ZTL may mediate homo- and hetero-dimerization of the three LOV/F-box/Kelch proteins and provide added insight into the composition of the protein complexes and an additional role for the Kelch repeat domain.

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Section: Ztl Kelch Repeat Interaction Profilessupporting

confidence: 63%

Section: Expression Of the Lov Domain Of Lov/f-box/kelch Proteins Dismentioning

confidence: 99%

Functional domain studies uncover novel roles for the ZTL Kelch repeat domain in clock function

Feke

Gendron

2020

Preprint

Self Cite

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“…2C), we engineered a dominant-negative (DN) form of SUSA2, where the F-box domain (residues 40-86) was deleted, likely abolishing the interaction of the F-box with SKP1/ASKs in the SCF E3 ligase complex but maintaining its interaction with its substrate, preventing self-ubiquitination and degradation of itself and its substrate. Such an approach has been used successfully for studying F-box proteins previously 32,33 . As expected, the DN-SUSA2-FLAG expressed well ( Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

SUSA2 is an F-box protein required for autoimmunity mediated by paired NLRs SOC3-CHS1 and SOC3-TN2

Liang

Tong

2020

Nat Commun

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Both higher plants and mammals rely on nucleotide-binding leucine-rich repeat (NLR) immune receptors to detect pathogens and initiate immunity. Upon effector recognition, plant NLRs oligomerize for defense activation, the mechanism of which is poorly understood. We previously showed that disruption of the E3 ligase, Senescence-Associated E3 Ubiquitin Ligase 1 (SAUL1) leads to the activation of the NLR SOC3. Here, we report the identification of suppressor of saul1 2 (susa2) and susa3 from the saul1-1 suppressor screen. Pairwise interaction analysis suggests that both SUSA proteins interact with components of an SCFSUSA2 E3 ligase complex as well as CHS1 or TN2, truncated NLRs that pair with SOC3. susa2-2 only suppresses the autoimmunity mediated by either CHS1 or TN2, suggesting its specific involvement in SOC3-mediated immunity. In summary, our study indicates links between plant NLRs and an SCF complex that may enable ubiquitination and degradation of unknown downstream components to activate defense.

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“…For example, deleting the F‐box domains of redundant F‐box proteins ZEITLUPE (ZTL), FLAVIN‐BINDING, KELCH REPEAT, F‐BOX1 (FKF1), or LOV KELCH PROTEIN2 (LKP2) leads to decoy versions of E3s, enabling genetic analysis of these redundant E3s and their substrates (Lee et al , ). Decoy versions of RING‐type E3s can be similarly generated by deleting the RING domain as in the case of redundant MAC3A (PUB59) and MAC3B (PUB60) (Feke et al , ).…”

Section: Discussionmentioning

confidence: 99%

Plant E3 ligases SNIPER 1 and SNIPER 2 broadly regulate the homeostasis of sensor NLR immune receptors

Tong

Tian

et al. 2020

The EMBO Journal

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In both plants and animals, nucleotide‐binding leucine‐rich repeat (NLR) immune receptors perceive pathogen‐derived molecules to trigger immunity. Global NLR homeostasis must be tightly controlled to ensure sufficient and timely immune output while avoiding aberrant activation, the mechanisms of which are largely unclear. In a previous reverse genetic screen, we identified two novel E3 ligases, SNIPER1 and its homolog SNIPER2, both of which broadly control the levels of NLR immune receptors in Arabidopsis. Protein levels of sensor NLRs (sNLRs) are inversely correlated with SNIPER1 amount and the interactions between SNIPER1 and sNLRs seem to be through the common nucleotide‐binding (NB) domains of sNLRs. In support, SNIPER1 can ubiquitinate the NB domains of multiple sNLRs in vitro. Our study thus reveals a novel process of global turnover of sNLRs by two master E3 ligases for immediate attenuation of immune output to effectively avoid autoimmunity. Such unique mechanism can be utilized in the future for engineering broad‐spectrum resistance in crops to fend off pathogens that damage our food supply.

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

Cited by 30 publications

References 120 publications

Functional domain studies uncover novel roles for the ZTL Kelch repeat domain in clock function

Functional domain studies uncover novel roles for the ZTL Kelch repeat domain in clock function

SUSA2 is an F-box protein required for autoimmunity mediated by paired NLRs SOC3-CHS1 and SOC3-TN2

Plant E3 ligases SNIPER 1 and SNIPER 2 broadly regulate the homeostasis of sensor NLR immune receptors

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