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Identification of FLS2 interacting proteins by co-immunoprecipitation and shot gun proteomics

ABSTRACT: To identify immediate early signaling components in the FLS2 pathway we performed co-immunoprecipitation (co-IP) experiments with FLS2-GFP as bait in Arabidopsis. We immunoprecipitated FLS2-GFP with anti-GFP antibody coated beads and analyzed co-precipitated proteins by liquid chromatography tandem mass spectrometry (LC-MS/MS).

INSTRUMENT(S): LTQ Orbitrap

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

TISSUE(S): Plant Cell, Seedling

SUBMITTER: Jan Sklenar

LAB HEAD: Frank Menke

PROVIDER: PXD003189 | Pride | 2016-01-26

REPOSITORIES: Pride

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			Action	DRS
	MES_C_1_1_120113 (F071470).mzid_MES_C_1_1_120113_(F071470).pride.mgf.gz	Mzid
	MES_C_1_1_120113 (F071470).pride.mztab.gz	Mztab
	MES_C_1_1_120113F071470.mzid.gz	Mzid
	MES_C_1_1_120113F071470.mzid_MES_C_1_1_120113_F071470.MGF	Mzid
	MES_C_1_1_1_120113.raw	Raw

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Publications

Attenuation of pattern recognition receptor signaling is mediated by a MAP kinase kinase kinase.

Mithoe Sharon C SC Ludwig Christina C Pel Michiel J C MJ Cucinotta Mara M Casartelli Alberto A Mbengue Malick M Sklenar Jan J Derbyshire Paul P Robatzek Silke S Pieterse Corné M J CM Aebersold Ruedi R Menke Frank L H FL

EMBO reports 20160114 3

Pattern recognition receptors (PRRs) play a key role in plant and animal innate immunity. PRR binding of their cognate ligand triggers a signaling network and activates an immune response. Activation of PRR signaling must be controlled prior to ligand binding to prevent spurious signaling and immune activation. Flagellin perception in Arabidopsis through FLAGELLIN-SENSITIVE 2 (FLS2) induces the activation of mitogen-activated protein kinases (MAPKs) and immunity. However, the precise molecular m ...[more]

PMID: 26769563

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Protein interaction partners of FRUITFULL in inflorescence meristems and pistils by IP-MS.

Project description:To study the possibility that the tissue-specific gene targets and regulation by FUL are due to a different composition in the FUL transcription factor (TF) protein complex/es in both tissues, we determined the protein-protein interactions of FUL in planta. Inflorescence meristems (IMs) of 35S:AP1-GR ap1 cal pFUL:FUL-GFP plants were collected to identify meristem-specific protein complexes of FUL, and stage 12 - 16 pistils of pFUL:FUL-GFP ful-1 plants were collected for the pistil-specific FUL protein complexes. Protein complexes were isolated by immunoprecipitation (IP) using anti-GFP antibodies and protein identification was performed using LC-MS/MS, followed by label-free protein quantification.

2023-03-30 | PXD037091 | Pride

Arabidopsis AGO1 N-terminal Poly-Q domain promotes phase separation and association with stress granules during heat stress

Project description:In Arabidopsis thaliana, ARGONAUTE1 (AGO1) plays a central role in microRNA (miRNA) and small interfering RNA (siRNA)-mediated silencing. Nuclear AGO1 is loaded with miRNAs and exported to the cytosol where it associates to the rough ER to conduct miRNA-mediated translational repression, mRNA cleavage and biogenesis of phased siRNAs. These latter, as well as other cytosolic siRNAs, are loaded into cytosolic AGO1, but in which compartment this happens is not known. Moreover, the effect of stress on AGO1 localization is still unclear. Here, we show that heat stress (HS) promotes AGO1 protein accumulation, which co-localize with components of the siRNA bodies and of stress granules (SGs). AGO1 does not need SGS3, a key component of siRNA bodies, to efficiently form condensates during HS. Instead, we found that the still poorly characterized N-terminal Poly-Q domain of AGO1, which contains a prion-like domain, is sufficient to undergo phase separation. Moreover, an exposure of 1 hour to HS only moderately affected AGO1 loading by miRNAs and target cleavage, suggesting that its localization in condensates protects AGO1 rather than promote its activity in reprograming gene expressing during stress. Collectively, our work shed new light on the impact of high temperature on a main effector of RNA silencing in plants.

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Murine Kidney Papilla Cells: Control GFP- vs. GFP+ label retaining cells. Cell populations were separated by FACS.

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LC-MS/MS analysis of isolated murine brain endothelial cells (BECs) from Arf6-GFP-AAV vs GFP-AAV treated WT mice followed by label-free quantification (LFQ)

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2024-02-15 | PXD045006 | Pride

Analysis of FAM65A interactions

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2016-11-01 | PXD004933 | Pride

Pexophagy suppresses ROS-induced damage in leaf cells under high-intensity light

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CALCOCO1 acts with VAMP-Associated Proteins to mediate ER-phagy

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