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The interaction proteins of FER were identified by mass spectrometry

ABSTRACT: Catharanthus roseus Receptor-Like Kinase 1-like (CrRLK1L) family proteins are considered as potential cell wall sensors in plants. CrRLK1L subfamily proteins harbor two extracellular malectin-like domains that may bind to cell wall polymers, and an intracellular kinase domain that transmits signals via the phosphorylation of intracellular substrates. In Arabidopsis, there are 17 CrRLK1L family members, among which FER is the most widely studied. A growing body of evidence support that FER not only regulates plant growth and development, such as root hair development, pavement cell morphogenesis, flowering, and fertilization, but also controls plant responses to biotic and abiotic stresses. To decipher molecular mechanisms underlying the FER-mediated regulation of plant growth and development, we performed an immunoprecipitation-mass spectrometry (IP-MS) assay using FER-GFP transgenic plants to search for FER-interacting proteins.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Plant Cell, Root, Shoot

SUBMITTER: Chunzhao Zhao

LAB HEAD: Chunzhao Zhao

PROVIDER: PXD047595 | Pride | 2024-08-30

REPOSITORIES: Pride

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			Action	DRS
	20200327_QP_ZCZ_1-CK.msf	Msf
	20200327_QP_ZCZ_1-CK.raw	Raw
	20200327_QP_ZCZ_2-FER-0h.msf	Msf
	20200327_QP_ZCZ_2-FER-0h.raw	Raw
	20200327_QP_ZCZ_3-FER-6h.msf	Msf

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The Identification of FERONIA‑interacting proteins in Arabidopsis thaliana via mass spectrometry

Project description:FERONIA (FER) is a plasma membrane-localized receptor-like kinase (RLK) that belongs to the Catharantus roseus RLK1-like (CrRLK1L) subfamily. FER serves as a potential cell wall sensor that regulates multiple phytohormones, including ABA, JA, SA, BR, auxin, and ethyl, but the underlying regulatory mechanisms are still largely unknown. To further understand how FER regulates downstream signaling pathway, we performed FER-interacting proteins via immunoprecipitation-mass spectrometry (IP-MS) assay generated from FER-GFP transgenic plants.

2024-09-14 | PXD047596 | Pride

FERONIA-mediated CC1 phosphorylation dynamically regulates cortical microtubule behavior and salt stress responses in Arabidopsis

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2024-08-30 | PXD047608 | Pride

FERONIA controls ABA-mediated seed germination via the regulation of the kinase activity of CARK1

Project description:Seed germination is the initial step of the whole life cycle for an individual plant, and thus it needs to be tightly controlled to avoid the growth of plants under unfavorable conditions, such as high salinity and drought stress. It has been well known that ABA signaling pathway plays a key role in the regulation of seed germination. In this study, we found that FER, a receptor-like kinase that belongs to CrRLK1L subfamily, regulates seed germination under ABA or stress conditions via the modulation of ABI5 protein stability. Biochemical data indicate that FER interacts with and phosphorylates CARK1 protein, a receptor-like cytoplasmic kinase (RLCK) that is classified into the RLCK VIII subfamily. FER phosphorylates the Ser233 and Thr234 residues of CARK1, and these two residues are located in the activation loop and are required for the activation of CARK1. The CARK1 protein with the substitutions of these two amino acids to Ala exhibits reduced kinase activity and is not able to rescue the faster seed germination phenotype of cark1 mutant under ABA conditions. In both fer-4 and cark1 mutant, the ABA-triggered activation of SnRK2.6 and ABI5 protein accumulation were attenuated compared with the wild-type. Taken together, our study not only reveals a RLCK protein that functions directly downstream of FER to transduce external signals, but also provides a novel mechanistic insight into the understanding of seed germination regulation via ABA-mediated signaling pathways.

2024-09-14 | PXD047594 | Pride

The LRX-RALF-FER cell wall integrity sensing module controls plant growth and salt stress responses by modulating multiple plant hormones

Project description:Plants have evolved cell wall integrity signaling pathways to maintain cell wall homeostasis during rapid growth and in response to environmental stress. The cell wall leucine-rich repeat extensins LRX3/4/5, the RAPID ALKALINIZATION FACTOR (RALF) peptides RALF22/23, and FERONIA (FER) function as a module to regulate plant growth and salt stress responses via the sense of cell wall integrity. However, the intracellular signaling pathways that mediate the effects of the LRX3/4/5-RALF22/23-FER module are still largely unknown. Here, we report that jasmonic acid (JA), salicylic acid (SA), and abscisic acid (ABA) accumulate constitutively in lrx345 and fer mutants. Blocking JA pathway rescues the retarded growth phenotype of the lrx345 and fer-4 mutants, while disruption of ABA biosynthesis suppresses the salt-hypersensitivity of these mutants. Many salt stress-responsive genes display abnormal expression patterns in the lrx345 and fer-4 mutants, as well as in wild type plants treated with epigallocatechin gallate (EGCG), an inhibitor of pectin methylesterases, suggesting that the cell wall integrity is a critical factor that determines the expression of stress-responsive genes. Production of reactive oxygen species (ROS) is constitutively increased in the lrx345 and fer-4 mutants, and inhibition of ROS accumulation suppresses the salt-hypersensitivity of these mutants. Together, our results suggest that the LRX3/4/5-RALF22/23-FER module controls plant growth and salt stress responses by regulating hormonal homeostasis and ROS accumulation.

2021-07-15 | GSE136268 | GEO

A regulatory module controls plant growth and stress response by modulating hormone level

Project description:Perception and relay of cell wall signals is critical for plants to regulate growth and stress responses, but the mechanism underlying this biological process remains largely unknown. Here we report that RAPID ALKALINIZATION FACTOR (RALFs) peptides, Leucine-rich repeat extensins (LRXs) and FERONIA (FER) are physically associated in extracellular region. lrx345, fer-4, and transgenic plants overexpressing RALF22 or RALF23 all showed retarded growth, increased sensitivity to salt stress, and constitutively increased levels of ABA, JA, and SA. Salt treatment or chemical disruption of cell wall promotes the release of mature RALF peptides, which negatively regulate the function of FER by inducing its internalization. JA mutants coi1 and aos restore the retarded growth, and mutation of ABA2 suppresses the salt-sensitive phenotype of lrx345. Together, we propose that LRXs, RALFs, and FER function as a module to sense and transduce cell wall signals, thereby regulating growth and stress responses via hormones-mediated pathways.

2021-07-15 | GSE114682 | GEO

Transcriptomic profile of FERONIA mutant treated with Aradopside

Project description:FERONIA (FER) receptor kinase has a versatile role in plant growth and development, biotic and abiotic stress responses, and reproduction. To gain new insights into the molecular interplay of these processes and to identify new functions of FER, we carried out quantitative transcriptome, proteome, and phosphoproteome profiling of wild type (WT) and a loss-of-function fer mutant in Arabidopsis plants. Gene Ontology terms for hormone signaling, abiotic stress, and biotic stress were significantly enriched among mis-expressed transcripts, proteins, and/or mis-phosphorylated proteins, in agreement with FER’s known roles in these processes. Analysis of multi-omics data and subsequent experimental evidence identified previously unknown functions of FER in ER (Endoplasmic Reticulum) body formation, glucosinolate biosynthesis. FER functions through transcription factor NAI1 to mediate ER body. FER also negatively regulates indole glucosinolates biosynthesis, partially through NAI1. Furthermore, we found that a group of abscisic acid (ABA)-induced transcription factors are hypo-phosphorylated in the fer mutant and demonstrated that FER acts through ABI5 to negatively regulate the ABA response during germination. Our integrated-omics study therefore reveals novel functions of FER and provides new insights into the underlying mechanisms of FER function.

2022-04-04 | GSE191303 | GEO

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Profiling FERONIA interaction proteins by Pupylation-based interacting tagging in Arabidopsis

Project description:FERONIA (FER) is a ubiquitious expressed and a plasma memberane-localized receptor-like protein kinase which modulate many biological functions. Elucidating the interacting proteins is the crucial step in understanding the molecular function of FER in regulating plant development. However, weak and transient interactions make identifying FER interacting proteins a challenge. Here, we adapted a novel proximity-based labelling technique, pupylation-based interaction tagging (PUP-IT), to label interacting proteins, subsequent enriched by affinity capture, and analyzed by liquid chromatography-coupled tandem mass spectrometery (LC-MS/MS) to profile FER interacting proteins. To perform a quick screen, a transient protoplasts expression system was conducted to identify interacting proteins in mesophyll cells. In addition, we generated a transgenic line that harboring a tagging substrate-inducible system and FER-ligase recombinant protein driven by a native promoter for surveying the interacting proteins in seedlings and flower organ. We introduce and prove the concept of PUP-IT tool designed to detect protein interactions of significance in plants.

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Profiling FERONIA interaction proteins by Pupylation-based interacting tagging in Arabidopsis

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