Protein Levels of Several Arabidopsis Auxin Response Factors Are Regulated by Multiple Factors and ABA Promotes ARF6 Protein Ubiquitination

Li, Keke; Wang, Sheng; Wu, Hong; Wang, Hong

doi:10.3390/ijms21249437

Cited by 25 publications

(18 citation statements)

References 64 publications

(99 reference statements)

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“…ARF7 and ARF19 are two class-A ARFs that are both regulated by the proteasome and directly interact with AFF1. Similarly, the class-A ARF6 13 , 14 and ARF8 13 , the Class-B ARF1 15 , ARF2 16 , and the Class-C ARF17 13 also undergo proteasome-dependent degradation. Whether these protein stabilities are regulated through the activity of SCF AFF1 or through another mechanism remains unknown.…”

Section: Discussionmentioning

confidence: 99%

Regulation of AUXIN RESPONSE FACTOR condensation and nucleo-cytoplasmic partitioning

et al. 2022

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Auxin critically regulates plant growth and development. Auxin-driven transcriptional responses are mediated through the AUXIN RESPONSE FACTOR (ARF) family of transcription factors. ARF protein condensation attenuates ARF activity, resulting in dramatic shifts in the auxin transcriptional landscape. Here, we perform a forward genetics screen for ARF hypercondensation, identifying an F-box protein, which we named AUXIN RESPONSE FACTOR F-BOX1 (AFF1). Functional characterization of SCFAFF1 revealed that this E3 ubiquitin ligase directly interacts with ARF19 and ARF7 to regulate their accumulation, condensation, and nucleo-cytoplasmic partitioning. Mutants defective in AFF1 display attenuated auxin responsiveness, and developmental defects, suggesting that SCFAFF1 -mediated regulation of ARF protein drives aspects of auxin response and plant development.

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Section: Discussionmentioning

confidence: 99%

Regulation of AUXIN RESPONSE FACTOR condensation and nucleo-cytoplasmic partitioning

et al. 2022

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“…Finally, the clearest example of ARF ubiquitination has been described for AtARF6. The mechanism is induced by abscisic acid and leads to the degradation of the protein (Li et al ., 2020).…”

Section: The Role Of Auxin Response Factor Domain‐mediated Interactio...mentioning

confidence: 99%

Auxin response factors are keys to the many auxin doors

et al. 2022

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Summary In plants, most developmental programs depend on the action of auxin. The best described model of the auxin signaling pathway, which explains most, but not all, of the auxin transcriptional responses, relies on a de‐repression mechanism. The auxin/indole‐3‐acetic acid repressors (Aux/IAAs) interact with the auxin response factors (ARFs), the transcription factors of the auxin signaling pathway, leading to repression of the ARF‐controlled genes. Auxin induces Aux/IAA degradation, releases ARFs and activates transcription. However, this elegant model is not suitable for all ARFs. Indeed, in Arabidopsis, which has 22 ARFs, only five of them fit into the model since they are the ones able to interact with Aux/IAAs. The remaining 17 have a limited capacity to interact with the repressors, and their mechanisms of action are still unclear. The differential interactions between ARF and Aux/IAA proteins constitute one of many examples of the biochemical and structural diversification of ARFs that affect their action and therefore affect auxin transcriptional responses. A deeper understanding of the structural properties of ARFs is fundamental to obtaining a better explanation of the action of auxin in plants.

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“…The HA-cyFBP protein was pulled down from the total protein extract as described [ 70 ]. The anti-HA agarose beads (Sigma-Aldrich) were added into the total protein extract (1 mg of total proteins) and incubated at 4 °C for 1 h (on a tumbler).…”

Section: Methodsmentioning

confidence: 99%

“…The bound proteins were eluted with 1 mg ml −1 HA peptide (GenScript, Piscataway, USA). After adding the 4X SDS loading buffer and heat treatment at 95 °C for 5 min, the samples were then analyzed by electrophoresis and Western blotting for HA-cyFBP and ubiquitinated HA-cyFBP as described [ 70 ].…”

Section: Methodsmentioning

confidence: 99%

Arabidopsis Ubiquitin-Conjugating Enzymes UBC4, UBC5, and UBC6 Have Major Functions in Sugar Metabolism and Leaf Senescence

Wang

Cao

Willick

et al. 2022

IJMS

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The ubiquitin-conjugating enzyme (E2) is required for protein ubiquitination. Arabidopsis has 37 E2s grouped into 14 subfamilies and the functions for many of them are unknown. We utilized genetic and biochemical methods to study the roles of Arabidopsis UBC4, UBC5, and UBC6 of the E2 subfamily IV. The Arabidopsis ubc4/5/6 triple mutant plants had higher levels of glucose, sucrose, and starch than the control plants, as well as a higher protein level of a key gluconeogenic enzyme, cytosolic fructose 1,6-bisphosphatase 1 (cyFBP). In an in vitro assay, the proteasome inhibitor MG132 inhibited the degradation of recombinant cyFBP whereas ATP promoted cyFBP degradation. In the quadruple mutant ubc4/5/6 cyfbp, the sugar levels returned to normal, suggesting that the increased sugar levels in the ubc4/5/6 mutant were due to an increased cyFBPase level. In addition, the ubc4/5/6 mutant plants showed early leaf senescence at late stages of plant development as well as accelerated leaf senescence using detached leaves. Further, the leaf senescence phenotype remained in the quadruple ubc4/5/6 cyfbp mutant. Our results suggest that UBC4/5/6 have two lines of important functions, in sugar metabolism through regulating the cyFBP protein level and in leaf senescence likely through a cyFBP-independent mechanism.

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Protein Levels of Several Arabidopsis Auxin Response Factors Are Regulated by Multiple Factors and ABA Promotes ARF6 Protein Ubiquitination

Cited by 25 publications

References 64 publications

Regulation of AUXIN RESPONSE FACTOR condensation and nucleo-cytoplasmic partitioning

Regulation of AUXIN RESPONSE FACTOR condensation and nucleo-cytoplasmic partitioning

Auxin response factors are keys to the many auxin doors

Arabidopsis Ubiquitin-Conjugating Enzymes UBC4, UBC5, and UBC6 Have Major Functions in Sugar Metabolism and Leaf Senescence

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