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Comparative proteomics of sink and source cell wall

ABSTRACT: Plants as multi cellular organisms are composed of various specialized cell types. Most of these cell types are symplasmically connected via plasmodesmata (PD) allowing exchange of nutrients and signal molecules. To date, the molecular composition of PD is not well understood. A comprehensive and comparative proteomic approach was followed to identify proteins providing new insights into the composition of these important regulatory structures.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Nicotiana Tabacum (common Tobacco)

TISSUE(S): Plant Cell, Leaf

SUBMITTER: Max Kraner

LAB HEAD: Uwe Sonnewald

PROVIDER: PXD007282 | Pride | 2017-09-06

REPOSITORIES: Pride

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			Action	DRS
	Sink_1_5ul.mgf	Mgf
	Sink_1_5ul.pride.mgf.gz	Mgf
	Sink_1_5ul.raw	Raw
	Sink_2_5ul.mgf	Mgf
	Sink_2_5ul.pride.mgf.gz	Mgf

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Publications

Comparative proteomic profiling of the choline transporter-like1 (CHER1) mutant provides insights into plasmodesmata composition of fully developed Arabidopsis thaliana leaves.

Kraner Max E ME Müller Carmen C Sonnewald Uwe U

The Plant journal : for cell and molecular biology 20171006 4

In plants, intercellular communication and exchange are highly dependent on cell wall bridging structures between adhering cells, so-called plasmodesmata (PD). In our previous genetic screen for PD-deficient Arabidopsis mutants, we described choline transporter-like 1 (CHER1) being important for PD genesis and maturation. Leaves of cher1 mutant plants have up to 10 times less PD, which do not develop to complex structures. Here we utilize the T-DNA insertion mutant cher1-4 and report a deep comp ...[more]

PMID: 28865150

Similar Datasets

Comparative proteomics of plasmodesmata

Project description:Plants as multi cellular organisms are composed of various specialized cell types. Most of these cell types are symplasmically connected via plasmodesmata (PD) allowing exchange of nutrients and signal molecules. To date, the molecular composition of PD is not well understood. A comprehensive and comparative proteomic approach was followed to identify proteins providing new insights into the composition of these important regulatory structures.

2017-09-05 | PXD006880 | Pride

Multiple C2 domains and Transmembrane region Proteins (MCTPs) tether membranes at plasmodesmata

Project description:In eukaryotes, membrane contact sites (MCS) allow direct communication between organelles. Plants have evolved unique MCS, the plasmodesmata intercellular pores, which combine organelle tethering with regulation of cell-to-cell signalling, the molecular mechanisms of which remains unknown. Here, we identify Multiple C2 domains and Transmembrane region Proteins (MCTPs) as tethers that link the endoplasmic reticulum (ER) to the plasma membrane (PM) within plasmodesmata. We report that MCTPs, including MCTP3 and 4 recently identified as modulators of SHOOTMERISTEMLESS trafficking, are ER-anchored proteins that cluster at plasmodesmata. MCTPs insert into the ER via their transmembrane region whilst their C2 domains dock to the PM through interaction with anionic phospholipids. A mctp3/4 loss-of function mutant induces plant developmental defects while MCTP4 expression in a yeast .tether mutant partially restores ER-PM tethering. Our data suggest that MCTPs are unique membrane tethers controlling both ER-PM contacts and cell-cell signalling.

2019-07-08 | PXD013999 | Pride

A high confidence Physcomitrium patens plasmodesmata proteome

Project description:Cells of multicellular organisms exchange nutrients, building blocks and information. In animals this exchange happens via gap junctions, in plants via plasmodesmata (PD). PD have striking properties with a large range of molecules from ions, to metabolites and synthetic dyes, RNA and proteins up to 40 kDa being translocated, ranging. PD are hard to characterize due to deep embedding into cell walls and presence of several membranes. Previous studies of the protein composition of PD of angiosperms identified large lists of proteins, few were validated. Here, we developed a probabilistic scoring approach in iteration with systematic localization on a large scale to define a high-confidence PD proteome of Physcomitrium patens. Conservation of targeting allowed us to validate 100s of proteins in tobacco. We developed a PD Score based on protein enrichment during PD preparation and significantly enriched protein features of known PD proteins from literature. PD scoring was iteratively improved by extensive validation of PD candidate protein localizations. As a result we establish a high confidence PD proteome from Physcomitrium patens comprising close to 300 proteins which together with the bona fide PD proteins from literature are made available in the public PDDB database. Conservation in localization strengthens the reliability of plant proteome and provides a basis for exploring the evolution of this important organelle. Glycolyl hydrolase family 17 proteins are presented as example of an abundant PD protein family with representatives across an evolutionary scale.

2024-01-18 | PXD032820 | Pride

Label-free quantitative proteomics on Nicotiana benthamiana plasmodesmal proteome following viral infection

Project description:We analyzed the PD-enriched fraction from Turnip mosaic virus (TuMV)-infected Nicotiana benthmiana (N. benthamiana) by using label-free quantitative proteomics of which 100 and 48 were significantly increased and decreased respectively when compared with mock plants.

2017-10-19 | PXD006323 | Pride

Comparative phyloproteomics identifies conserved plasmodesmal proteins

Project description:Plasmodesmata are cytosolic bridges, lined by the plasma membrane and traversed by endoplasmic reticulum, connecting cells and tissues, and critical for many aspects of plant biology. While plasmodesmata are notoriously difficult to extract, tissue fractionation and proteomic analyses can yield valuable knowledge of their composition. Here we have generated two novel proteomes to expand tissue and taxonomic representation of plasmodesmata: one from mature Arabidopsis leaves and one from the moss Physcomitrium patens and leveraged these and existing data to perform a comparative analysis to identify evolutionarily conserved protein families that are associated with plasmodesmata. Thus, we identified b-1,3-glucanases, C2 lipid-binding proteins and tetraspanins as core plasmodesmal components that likely serve as essential structural or functional components. Our approach has not only identified elements of a conserved plasmodesmal proteome, but also demonstrated the added power offered by comparative analysis for recalcitrant samples. Conserved plasmodesmal proteins establish a basis upon which ancient plasmodesmal function can be further investigated to determine the essential roles these structures play in multicellular organism physiology in the green lineages.

2023-01-13 | PXD038964 | Pride

Choline transporter-like1 is required for plasmodesmal targeting of Potato leafroll virus movement protein MP17 and long-distance assimilate transport

Project description:Macromolecular trafficking and cell-to-cell communication in plants occurs via plasmodesmata (PD). Currently, little is known about the proteins defining these sophisticated cell-to-cell contacts. To uncover proteins required for PD structure and function we made use of the 17 kDa movement protein (MP17) of the Potato leafroll virus (PLRV). The protein is required for cell-to-cell movement of the virus and localizes specifically to branched PD in source tissues. By forward genetic screening for Arabidopsis mutants with altered PD binding of MP17, several mutants were found. Map-based cloning of one of these mutants revealed a mutation in the choline transporter-like 1 (CHER1) protein. The mutation abolished plasma membrane localization of the protein. As consequence phosphatidylcholine levels and PD binding of MP17 decreased. Transcriptome analysis revealed a down-regulation of defense genes and genes involved in the synthesis of very long chain fatty acids, indicating an impairment of lipid signaling. Furthermore, cher1 mutants showed a reduced assimilate export and stunted growth. These findings highlight the emerging role of phospholipids, especially in the context of PD structure and function as well as potential binding sites for plasma membrane- and PD-associated proteins, which has been neglected for a long time. light exposed source leaves from wild type Col-0, T-DNA insertion line cher1-4 and mutant cher1-5 (10 weeks old). Samples: per pool 2 source leaves from 4 different plants, taken in the afternoon; wildtype (WT): Col-0, cher1-4: T-DNA insertion line SALK-065853 (ecotype Col-0, kanamycin resistance), cher1-5: Col-0 with GGA -->GAA at position 740 of cds

2016-06-01 | E-GEOD-61595 | biostudies-arrayexpress

Expression profiles of embryo-defective Arabidopsis thaliana mutants increased size exclusion limit1 (ise1) and ise2

Project description:We use Arabidopsis thaliana embryogenesis as a model system for studying intercellular transport via plasmodesmata (PD). A forward genetic screen for altered PD transport identified ise1 and ise2 mutants with increased intercellular transport of fluorescent 10-kDa tracers. Both ise1 and ise2 exhibit increased formation of twinned and branched PD. ISE1 encodes a mitochondrial DEAD-box RNA helicase, while ISE2 encodes a DEVH-type RNA helicase. Here we show that ISE2 foci are localized to the chloroplast stroma. Surprisingly, plastid development is defective in both ise1 and ise2 mutant embryos. In an effort to understand how RNA helicases that localize to different organelles have similar impacts on plastid and PD development/function we performed whole genome expression analyses. The most significantly affected class of transcripts in both mutants encodes products that target to and enable plastid function. These results reinforce the importance of plastid-mitochondria-nucleus crosstalk, add PD as a critical player in the plant cell communication network, and thereby illuminate a new signaling pathway, dubbed organelle-nucleus-plasmodesmata signaling (ONPS). Several genes with roles in cell wall synthesis and modification are also differentially expressed in both mutants, providing new targets for investigating PD development and function. Three biological replicates each of either ise1 or ise2 mutant seeds vs. sister wild-type controls

2011-11-09 | E-GEOD-33558 | biostudies-arrayexpress

Comparison of gene expression profiles of Jurkat cells with or without PD-1 ligation

Project description:CD8+ T cells in chronic viral infections like HIV develop functional defects such as loss of IL-2 secretion and decreased proliferative potential that are collectively termed exhaustion1. Exhausted T cells express increased levels of multiple inhibitory receptors, such as Programmed Death 1 (PD-1). PD-1 inhibition contributes to impaired virus-specific T cell function in chronic infection because antibody-mediated blockade of its ligand, Programmed Death Ligand 1 (PD-L1) is sufficient to improve T cell function and reduce viral replication in animal models. Reversing PD-1 inhibition is therefore an attractive therapeutic target, but the cellular mechanisms by which PD-1 ligation results in T cell inhibition are not fully understood. PD-1 is thought to limit T cell activation by attenuating T cell receptor (TCR) signaling. It is not known whether PD-1 ligation also acts by upregulating genes in exhausted T cells that impair their function. Here, we analyzed gene-expression profiles from HIV-specific CD8+ T cells in patients with HIV and show that PD-1 coordinately upregulates a program of genes in exhausted CD8+ T cells from humans and mice. This program includes upregulation of basic leucine transcription factor, ATF-like (BATF), a transcription factor in the AP-1 family. Enforced expression of BATF was sufficient to impair T cell proliferation and cytokine secretion, while BATF knockdown reduced PD-1 inhibition. Silencing BATF in CD4+ and CD8+ T cells from chronic viremic patients rescued HIV-specific T cell function. Thus inhibitory receptors can cause T cell exhaustion by upregulating genes â such as BATF â that inhibit T cell function. PD-1 expressing Jurkat cells were cultured for 18 hours with beads coated with antibodies to CD3 and CD28, with our without an antibody to PD-1.

2010-10-03 | E-GEOD-24026 | biostudies-arrayexpress

Gene expression profile of mouse breast cancer V720 cells treated with vehicle or PD 0332991

Project description:D-cyclins represent components of cell cycle machinery. To test the efficacy of targeting D-cyclins in cancer treatment, we engineered mouse strains which allow acute and global ablation of individual D-cyclins in a living animal. Ubiquitous shutdown of cyclin D1 or inhibition of cyclin D associated kinase activity in mice bearing ErbB2-driven mammary carcinomas halted cancer progression and triggered tumor-specific senescence, without compromising the animals' health. Ablation of cyclin D3 in mice bearing T-cell acute lymphoblastic leukemias (T-ALL) triggered tumorspecific apoptosis. Such selective killing of leukemic cells can be also achieved by inhibiting cyclin D associated kinase activity in mouse and human T-ALL models. Hence, contrary to what one might expect from ablation of a cell cycle protein, acute shutdown of a D-cyclin leads not only to cell cycle arrest, but it also triggers tumor cell senescence or apoptosis, and it affects different tumor types through distinct cellular mechanisms. Inhibiting cyclin D-activity represents a highly-selective anticancer strategy which specifically targets cancer cells without significantly affecting normal tissues. Mouse breast cancer V720 cells were cultured in the presence of the CDK4/6 inhibitor PD 0332991 (PD; 1 microM) or vehicle (VO) for 24 hrs. Experiment was done in biological triplicate. A total of 6 RNA samples (3 vehicle treated and 3 PD 0332991 treated samples) were used for microarray expression analysis.

2012-10-16 | E-GEOD-40513 | biostudies-arrayexpress

The Requirement for Cyclin D Function in Tumor Maintenance

Project description:This SuperSeries is composed of the following subset Series: GSE40512: Gene expression profile of human T-ALL cell line KOPTK1 treated with vehicle or PD 0332991 GSE40513: Gene expression profile of mouse breast cancer V720 cells treated with vehicle or PD 0332991 Refer to individual Series

2012-10-16 | E-GEOD-40514 | biostudies-arrayexpress

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