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1.
Plant Physiol ; 189(2): 557-566, 2022 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-35099565

RESUMO

The exocyst is the main plasma membrane vesicle-tethering complex in eukaryotes and is composed of eight different subunits. Yet, in plant genomes, many subunits display multiple copies, thought to reflect evolution of complex subtypes with divergent functions. In Arabidopsis thaliana root endodermal cells, the isoform EXO70A1 is required for positioning of CASP1 at the Casparian Strip Domain, but not for its non-targeted secretion to the plasma membrane. Here, we show that exo84b resembles exo70a1 mutants regarding CASP1 mistargeting and secretion of apoplastic proteins, but exo84b additionally affects secretion of other integral plasma membrane proteins. Moreover, conditional, cell-type-specific gene editing of the single-copy core component SEC6 allows visualization of secretion defects in plant cells with a complete lack of exocyst complex function. Our approach opens avenues for deciphering the complexity/diversity of exocyst functions in plant cells and enables analysis of central trafficking components with lethal phenotypes.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Membrana Celular/metabolismo , Parede Celular/metabolismo , Citoplasma/metabolismo , Proteínas de Membrana/metabolismo
2.
Proc Natl Acad Sci U S A ; 117(46): 29166-29177, 2020 11 17.
Artigo em Inglês | MEDLINE | ID: mdl-33139576

RESUMO

Lignin has enabled plants to colonize land, grow tall, transport water within their bodies, and protect themselves against various stresses. Consequently, this polyphenolic polymer, impregnating cellulosic plant cell walls, is the second most abundant polymer on Earth. Yet, despite its great physiological, ecological, and economical importance, our knowledge of lignin biosynthesis in vivo, especially the polymerization steps within the cell wall, remains vague-specifically, the respective roles of the two polymerizing enzymes classes, laccases and peroxidases. One reason for this lies in the very high numbers of laccases and peroxidases encoded by 17 and 73 homologous genes, respectively, in Arabidopsis Here, we have focused on a specific lignin structure, the ring-like Casparian strips (CSs) within the root endodermis. By reducing candidate numbers using cellular resolution expression and localization data and by boosting stacking of mutants using CRISPR-Cas9, we mutated the majority of laccases in Arabidopsis in a nonuple mutant-essentially abolishing laccases with detectable endodermal expression. Yet, we were unable to detect even slight defects in CS formation. By contrast, we were able to induce a complete absence of CS formation in a quintuple peroxidase mutant. Our findings are in stark contrast to the strong requirement of xylem vessels for laccase action and indicate that lignin in different cell types can be polymerized in very distinct ways. We speculate that cells lignify differently depending on whether lignin is localized or ubiquitous and whether cells stay alive during and after lignification, as well as the composition of the cell wall.


Assuntos
Lacase/genética , Lacase/metabolismo , Peroxidases/genética , Peroxidases/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Membrana Celular/metabolismo , Parede Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Lignina/metabolismo , Mutação , Fenótipo , Raízes de Plantas , Polimerização , Xilema/metabolismo
3.
Plant Physiol ; 182(4): 1920-1932, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31992602

RESUMO

Phytochelatin synthase (PCS) is a key component of heavy metal detoxification in plants. PCS catalyzes both the synthesis of the peptide phytochelatin from glutathione and the degradation of glutathione conjugates via peptidase activity. Here, we describe a role for PCS in disease resistance against plant pathogenic fungi. The pen4 mutant, which is allelic to cadmium insensitive1 (cad1/pcs1) mutants, was recovered from a screen for Arabidopsis mutants with reduced resistance to the nonadapted barley fungal pathogen Blumeria graminis f. sp. hordei PCS1, which is found in the cytoplasm of cells of healthy plants, translocates upon pathogen attack and colocalizes with the PEN2 myrosinase on the surface of immobilized mitochondria. pcs1 and pen2 mutant plants exhibit similar metabolic defects in the accumulation of pathogen-inducible indole glucosinolate-derived compounds, suggesting that PEN2 and PCS1 act in the same metabolic pathway. The function of PCS1 in this pathway is independent of phytochelatin synthesis and deglycination of glutathione conjugates, as catalytic-site mutants of PCS1 are still functional in indole glucosinolate metabolism. In uncovering a peptidase-independent function for PCS1, we reveal this enzyme to be a moonlighting protein important for plant responses to both biotic and abiotic stresses.


Assuntos
Ascomicetos/metabolismo , Mitocôndrias/metabolismo , Fitoquelatinas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Catálise , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia
4.
Plant Physiol ; 179(4): 1581-1593, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30718348

RESUMO

Physiological acclimation of plants to an everchanging environment is governed by complex combinatorial signaling networks that perceive and transduce various abiotic and biotic stimuli. Reactive oxygen species (ROS) serve as one of the second messengers in plant responses to hyperosmotic stress. The molecular bases of ROS production and the primary cellular processes that they target were investigated in the Arabidopsis (Arabidopsis thaliana) root. Combined pharmacological and genetic approaches showed that the RESPIRATORY BURST OXIDASE HOMOLOG (RBOH) pathway and an additional pathway involving apoplastic ascorbate and iron can account for ROS production upon hyperosmotic stimulation. The two pathways determine synergistically the rate of membrane internalization, within minutes after activation. Live superresolution microscopy revealed at single-molecule scale how ROS control specific diffusion and nano-organization of membrane cargo proteins. In particular, ROS generated by RBOHs initiated clustering of the PLASMA MEMBRANE INTRINSIC PROTEIN2;1 aquaporin and its removal from the plasma membrane. This process is contributed to by clathrin-mediated endocytosis, with a positive role of RBOH-dependent ROS, specifically under hyperosmotic stress.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Pressão Osmótica , Espécies Reativas de Oxigênio/metabolismo , Aquaporinas/análise , Aquaporinas/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/análise , Proteínas de Arabidopsis/química , Endocitose , Domínios Proteicos , Transdução de Sinais
5.
PLoS Genet ; 12(2): e1005817, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26828932

RESUMO

Correct gene expression requires tight RNA quality control both at transcriptional and post-transcriptional levels. Using a splicing-defective allele of PASTICCINO2 (PAS2), a gene essential for plant development, we isolated suppressor mutations modifying pas2-1 mRNA profiles and restoring wild-type growth. Three suppressor of pas2 (sop) mutations modified the degradation of mis-spliced pas2-1 mRNA species, allowing the synthesis of a functional protein. Cloning of the suppressor mutations identified the core subunit of the exosome SOP2/RRP4, the exosome nucleoplasmic cofactor SOP3/HEN2 and a novel zinc-finger protein SOP1 that colocalizes with HEN2 in nucleoplasmic foci. The three SOP proteins counteract post-transcriptional (trans)gene silencing (PTGS), which suggests that they all act in RNA quality control. In addition, sop1 mutants accumulate some, but not all of the misprocessed mRNAs and other types of RNAs that are observed in exosome mutants. Taken together, our data show that SOP1 is a new component of nuclear RNA surveillance that is required for the degradation of a specific subset of nuclear exosome targets.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Transporte/metabolismo , Núcleo Celular/metabolismo , Exossomos/metabolismo , Dedos de Zinco , Alelos , Processamento Alternativo/genética , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Transporte/genética , Genes Supressores , Loci Gênicos , Íntrons/genética , Mutação/genética , Degradação do RNAm Mediada por Códon sem Sentido , Proteínas Nucleares/metabolismo , Isoformas de Proteínas/metabolismo , Processamento Pós-Transcricional do RNA/genética , Sítios de Splice de RNA/genética
6.
J Exp Bot ; 68(16): 4583-4593, 2017 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-28981771

RESUMO

Perturbation of cellulose synthesis in plants triggers stress responses, including growth retardation, mediated by the cell wall integrity-sensing receptor-like kinase (RLK) THESEUS1 (THE1). The analysis of two alleles carrying T-DNA insertions at comparable positions has led to conflicting conclusions concerning the impact of THE1 signaling on growth. Here we confirm that, unlike the1-3 and other the1 alleles in which cellular responses to genetic or pharmacological inhibition of cellulose synthesis are attenuated, the1-4 showed enhanced responses, including growth inhibition, ectopic lignification, and stress gene expression. Both the1-3 and the1-4 express a transcript encoding a predicted membrane-associated truncated protein lacking the kinase domain. However, the1-3, in contrast to the1-4, strongly expresses antisense transcripts, which are expected to prevent the expression of the truncated protein as suggested by the genetic interactions between the two alleles. Seedlings overexpressing such a truncated protein react to isoxaben treatment similarly to the1-4 and the full-length THE overexpressor. We conclude that the1-4 is a hypermorphic allele; that THE1 signaling upon cell wall damage has a negative impact on cell expansion; and that caution is required when interpreting the phenotypic effects of T-DNA insertions in RLK genes.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/citologia , Parede Celular/metabolismo , Proteínas Quinases/genética , Receptores de Superfície Celular/genética , Alelos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Benzamidas/farmacologia , Parede Celular/genética , Celulose/biossíntese , DNA Bacteriano , Regulação da Expressão Gênica de Plantas , Genes Dominantes , Lignina/metabolismo , Plantas Geneticamente Modificadas , Proteínas Quinases/metabolismo , Receptores de Superfície Celular/metabolismo , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/metabolismo , Transdução de Sinais
7.
J Biol Chem ; 289(50): 34911-20, 2014 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-25331944

RESUMO

Mutations in the Arabidopsis COBRA gene lead to defects in cellulose synthesis but the function of COBRA is unknown. Here we present evidence that COBRA localizes to discrete particles in the plasma membrane and is sensitive to inhibitors of cellulose synthesis, suggesting that COBRA and the cellulose synthase complex reside in close proximity on the plasma membrane. Live-cell imaging of cellulose synthesis indicated that, once initiated, cellulose synthesis appeared to proceed normally in the cobra mutant. Using isothermal calorimetry, COBRA was found to bind individual ß1-4-linked glucan chains with a KD of 3.2 µm. Competition assays suggests that COBRA binds individual ß1-4-linked glucan chains with higher affinity than crystalline cellulose. Solid-state nuclear magnetic resonance studies of the cell wall of the cobra mutant also indicated that, in addition to decreases in cellulose amount, the properties of the cellulose fibrils and other cell wall polymers differed from wild type by being less crystalline and having an increased number of reducing ends. We interpret the available evidence as suggesting that COBRA facilitates cellulose crystallization from the emerging ß1-4-glucan chains by acting as a "polysaccharide chaperone."


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Arabidopsis/metabolismo , Membrana Celular/metabolismo , Celulose/química , Glicoproteínas de Membrana/metabolismo , Parede Celular/metabolismo , Cristalização , Glucanos/química , Glucanos/metabolismo , Imagem Molecular , Transporte Proteico
8.
Plant J ; 78(1): 121-33, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24479634

RESUMO

Growth is a complex trait that adapts to the prevailing conditions by integrating many internal and external signals. Understanding the molecular origin of this variation remains a challenging issue. In this study, natural variation of shoot growth under mannitol-induced stress was analyzed by standard quantitative trait locus mapping methods in a recombinant inbred line population derived from a cross between the Col-0 and Cvi-0 Arabidopsis thaliana accessions. Cloning of a major QTL specific to mannitol-induced stress condition led to identification of EGM1 and EGM2, a pair of tandem-duplicated genes encoding receptor-like kinases that are potentially involved in signaling of mannitol-associated stress responses. Using various genetic approaches, we identified two non-synonymous mutations in the EGM2[Cvi] allele that are shared by at least ten accessions from various origins and are probably responsible for a specific tolerance to mannitol. We have shown that the enhanced shoot growth phenotype contributed by the Cvi allele is not linked to generic osmotic properties but instead to a specific chemical property of mannitol itself. This result raises the question of the function of such a gene in A. thaliana, a species that does not synthesize mannitol. Our findings suggest that the receptor-like kinases encoded by EGM genes may be activated by mannitol produced by pathogens such as fungi, and may contribute to plant defense responses whenever mannitol is present.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Manitol/farmacologia , Estresse Fisiológico , Alelos , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Mapeamento Cromossômico , Variação Genética , Interações Hospedeiro-Patógeno , Mutação , Fenótipo , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/enzimologia , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Locos de Características Quantitativas
9.
Biochim Biophys Acta ; 1842(10): 1422-30, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25004376

RESUMO

Plant cytokinesis requires intense membrane trafficking and remodeling to form a specific membrane structure, the cell plate that will ultimately separate the daughter cells. The nature and the role of lipids involved in the formation of the cell plate remain unclear. Plant membranes are particularly rich in sphingolipids such as glucosyl-ceramides with long (16 carbons) or very long (24 carbons) acyl chains. We reveal here that inhibition of the synthesis of sphingolipids with very long acyl chains induces defective cell plates with persistent vesicular structures and large gaps. Golgi-derived vesicles carrying material toward the cell plate display longer vesicle-vesicle contact time and their cargos accumulate at the cell plate, suggesting membrane fusion and/or recycling defects. In vitro fusion experiments between artificial vesicles show that glycosphingolipids with very long acyl chains stimulate lipid bilayer fusion. Therefore we propose that the very long acyl chains of sphingolipids are essential structural determinants for vesicle dynamics and membrane fusion during cytokinesis.

10.
Plant Cell ; 24(2): 589-607, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22327741

RESUMO

Plant cells are encased by a cellulose-containing wall that is essential for plant morphogenesis. Cellulose consists of ß-1,4-linked glucan chains assembled into paracrystalline microfibrils that are synthesized by plasma membrane-located cellulose synthase (CESA) complexes. Associations with hemicelluloses are important for microfibril spacing and for maintaining cell wall tensile strength. Several components associated with cellulose synthesis have been identified; however, the biological functions for many of them remain elusive. We show that the chitinase-like (CTL) proteins, CTL1/POM1 and CTL2, are functionally equivalent, affect cellulose biosynthesis, and are likely to play a key role in establishing interactions between cellulose microfibrils and hemicelluloses. CTL1/POM1 coincided with CESAs in the endomembrane system and was secreted to the apoplast. The movement of CESAs was compromised in ctl1/pom1 mutant seedlings, and the cellulose content and xyloglucan structures were altered. X-ray analysis revealed reduced crystalline cellulose content in ctl1 ctl2 double mutants, suggesting that the CTLs cooperatively affect assembly of the glucan chains, which may affect interactions between hemicelluloses and cellulose. Consistent with this hypothesis, both CTLs bound glucan-based polymers in vitro. We propose that the apoplastic CTLs regulate cellulose assembly and interaction with hemicelluloses via binding to emerging cellulose microfibrils.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Celulose/biossíntese , Quitinases/metabolismo , Glucanos/metabolismo , Glicosídeo Hidrolases/metabolismo , Arabidopsis/enzimologia , Proteínas de Arabidopsis/genética , Parede Celular/metabolismo , Quitinases/genética , Glicosídeo Hidrolases/genética , Microfibrilas/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Polissacarídeos/metabolismo
11.
Plant Cell ; 23(6): 2362-78, 2011 06.
Artigo em Inglês | MEDLINE | ID: mdl-21666002

RESUMO

Sphingolipids are a class of structural membrane lipids involved in membrane trafficking and cell polarity. Functional analysis of the ceramide synthase family in Arabidopsis thaliana demonstrates the existence of two activities selective for the length of the acyl chains. Very-long-acyl-chain (C > 18 carbons) but not long-chain sphingolipids are essential for plant development. Reduction of very-long-chain fatty acid sphingolipid levels leads in particular to auxin-dependent inhibition of lateral root emergence that is associated with selective aggregation of the plasma membrane auxin carriers AUX1 and PIN1 in the cytosol. Defective targeting of polar auxin carriers is characterized by specific aggregation of Rab-A2(a)- and Rab-A1(e)-labeled early endosomes along the secretory pathway. These aggregates correlate with the accumulation of membrane structures and vesicle fragmentation in the cytosol. In conclusion, sphingolipids with very long acyl chains define a trafficking pathway with specific endomembrane compartments and polar auxin transport protein cargoes.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Membrana Celular/metabolismo , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Via Secretória/fisiologia , Esfingolipídeos , Sequência de Aminoácidos , Animais , Arabidopsis/citologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Brefeldina A/metabolismo , Polaridade Celular , Ceramidas/química , Ceramidas/metabolismo , Endossomos/metabolismo , Inibidores Enzimáticos/metabolismo , Fumonisinas/metabolismo , Humanos , Ácidos Indolacéticos/metabolismo , Membranas Intracelulares/metabolismo , Membranas Intracelulares/ultraestrutura , Isoenzimas/genética , Isoenzimas/metabolismo , Proteínas de Membrana/genética , Dados de Sequência Molecular , Oxirredutases/antagonistas & inibidores , Oxirredutases/genética , Oxirredutases/metabolismo , Inibidores da Síntese de Proteínas/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Alinhamento de Sequência , Esfingolipídeos/química , Esfingolipídeos/metabolismo
13.
Nat Commun ; 14(1): 1626, 2023 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-36959183

RESUMO

Casparian strips (CS) are aligned bands of lignin-impregnated cell walls, building an extracellular diffusion barrier in roots. Their structure profoundly differs from tight junctions (TJ), analogous structures in animals. Nonetheless, CS membrane domain (CSD) proteins 1-5 (CASP1-5) are homologues of occludins, TJ components. CASP-marked membranes display cell wall (matrix) adhesion and membrane protein exclusion. A full CASP knock-out now reveals CASPs are not needed for localized lignification, since correctly positioned lignin microdomains still form in the mutant. Ultra-structurally, however, these microdomains are disorganized, showing excessive cell wall growth, lack of exclusion zone and matrix adhesion, and impaired exocyst dynamics. Proximity-labelling identifies a Rab-GTPase subfamily, known exocyst activators, as potential CASP-interactors and demonstrate their localization and function at the CSD. We propose that CASP microdomains displace initial secretory foci by excluding vesicle tethering factors, thereby ensuring rapid fusion of microdomains into a membrane-cell wall band that seals the extracellular space.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Lignina/metabolismo , Membrana Celular/metabolismo , Transporte Biológico
14.
Curr Opin Plant Biol ; 11(3): 321-8, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18395489

RESUMO

The growth of plant cells involves a constant adjustment of synthesis and rearrangement of cell wall polymers. Recently, three plasma membrane-bound receptor kinases related to CrRLK1 have been shown to be involved in the negative control of cell growth in different contexts. THESEUS1 is activated in mutants deficient for cellulose and may act as a cell wall integrity sensor inhibiting cell elongation. FERONIA is polarly localized in synergid cells of the female gametophyte and is required for growth cessation of compatible pollen tubes and subsequent delivery of sperm cells. AmRLK is involved in the control of the polar conical outgrowth of epidermal cells of Antirrhinum petals. The conservation of both extracellular and kinase domains suggests that the three receptors bind to related ligands and have similar cellular outputs, which may involve the production of reactive oxygen species.


Assuntos
Proteínas de Plantas/metabolismo , Plantas/enzimologia , Proteínas Quinases/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiologia , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Fosfotransferases/genética , Fosfotransferases/metabolismo , Fosfotransferases/fisiologia , Desenvolvimento Vegetal , Proteínas de Plantas/genética , Plantas/genética , Proteínas Quinases/genética , Proteínas Quinases/fisiologia , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Receptores de Superfície Celular/fisiologia
15.
Curr Biol ; 17(11): 922-31, 2007 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-17540573

RESUMO

BACKGROUND: A major challenge is to understand how the walls of expanding plant cells are correctly assembled and remodeled, often in the presence of wall-degrading micro-organisms. Plant cells, like yeast, react to cell-wall perturbations as shown by changes in gene expression, accumulation of ectopic lignin, and growth arrest caused by the inhibition of cellulose synthesis. RESULTS: We have identified a plasma-membrane-bound receptor-like kinase (THESEUS1), which is present in elongating cells. Mutations in THE1 and overexpression of a functional THE1-GFP fusion protein did not affect wild-type (WT) plants but respectively attenuated and enhanced growth inhibition and ectopic lignification in seedlings mutated in cellulose synthase CESA6 without influencing the cellulose deficiency. A T-DNA insertion mutant for THE1 also attenuated the growth defect and ectopic-lignin production in other but not all cellulose-deficient mutants. The deregulation of a small number of genes in cesA6 mutants depended on the presence of THE1. Some of these genes are involved in pathogen defense, in wall crosslinking, or in protecting the cell against reactive oxygen species. CONCLUSIONS: The results show that THE1 mediates the response of growing plant cells to the perturbation of cellulose synthesis and may act as a cell-wall-integrity sensor.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/enzimologia , Celulose/biossíntese , Proteínas Quinases/fisiologia , Receptores de Superfície Celular/fisiologia , Alelos , Sequência de Aminoácidos , Arabidopsis/citologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Parede Celular/metabolismo , Citoesqueleto/metabolismo , Citoesqueleto/ultraestrutura , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Proteínas de Fluorescência Verde/análise , Hipocótilo/enzimologia , Hipocótilo/crescimento & desenvolvimento , Hipocótilo/metabolismo , Lignina/metabolismo , Dados de Sequência Molecular , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo
16.
Curr Biol ; 28(15): 2452-2458.e4, 2018 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-30057301

RESUMO

The growth of plants, like that of other walled organisms, depends on the ability of the cell wall to yield without losing its integrity. In this context, plant cells can sense the perturbation of their walls and trigger adaptive modifications in cell wall polymer interactions. Catharanthus roseus receptor-like kinase 1-like (CrRLK1L) THESEUS1 (THE1) was previously shown in Arabidopsis to trigger growth inhibition and defense responses upon perturbation of the cell wall, but so far, neither the ligand nor the role of the receptor in normal development was known. Here, we report that THE1 is a receptor for the peptide rapid alkalinization factor (RALF) 34 and that this signaling module has a role in the fine-tuning of lateral root initiation. We also show that RALF34-THE1 signaling depends, at least for some responses, on FERONIA (FER), another RALF receptor involved in a variety of processes, including immune signaling, mechanosensing, and reproduction [1]. Together, the results show that RALF34 and THE1 are part of a signaling network that integrates information on the integrity of the cell wall with the coordination of normal morphogenesis.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Hormônios Peptídicos/genética , Raízes de Plantas/crescimento & desenvolvimento , Proteínas Quinases/genética , Receptores de Superfície Celular/genética , Transdução de Sinais , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Parede Celular/metabolismo , Hormônios Peptídicos/metabolismo , Raízes de Plantas/genética , Proteínas Quinases/metabolismo , Receptores de Superfície Celular/metabolismo
17.
Nat Plants ; 3: 17058, 2017 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-28436943

RESUMO

In a striking case of evolutionary convergence, polarized cell layers with ring-like diffusion barriers have evolved in both plant and animal lineages independently. In plants, ring-like Casparian strips become localized by the CASPARIAN STRIP MEMBRANE DOMAIN PROTEINS (CASPs). The mechanism of this striking localization, however, has remained enigmatic. Here we present a genetic screen aimed at isolating determinants of CASP localization. One of the mutants, lord of the rings 2 (lotr2)/exo70a1, displays dramatic de-localization of CASPs into randomly localized microdomains. EXO70A1 is a subunit of the exocyst complex, a central component of secretion in eukaryotes. Irradiation of EXO70 subunit genes in plants has suggested specialization of this conserved complex. Intriguingly, lotr2/exo70a1 does neither affect secretion of the CASPs, nor that of other membrane proteins in the endodermis, thus separating exocyst activity in localization from a general defect in secretion. Our results establish EXO70A1 as a central player in Casparian strip formation, generating a transient positional information that will be translated into a precisely localized cell wall modification.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Parede Celular/metabolismo , Proteínas de Membrana/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Membrana/metabolismo
18.
PLoS One ; 11(9): e0160631, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27583779

RESUMO

Very long chain fatty acids (VLCFAs) are involved in plant development and particularly in several cellular processes such as membrane trafficking, cell division and cell differentiation. However, the precise role of VLCFAs in these different cellular processes is still poorly understood in plants. In order to identify new factors associated with the biosynthesis or function of VLCFAs, a yeast multicopy suppressor screen was carried out in a yeast mutant strain defective for fatty acid elongation. Loss of function of the elongase 3 hydroxyacyl-CoA dehydratase PHS1 in yeast and PASTICCINO2 in plants prevents growth and induces cytokinesis defects. PROTEIN TYROSIN PHOSPHATASE-LIKE (PTPLA) previously characterized as an inactive dehydratase was able to restore yeast phs1 growth and VLCFAs elongation but not the plant pas2-1 defects. PTPLA interacted with elongase subunits in the Endoplasmic Reticulum (ER) and its absence induced the accumulation of 3-hydroxyacyl-CoA as expected from a dehydratase involved in fatty acid (FA) elongation. However, loss of PTPLA function increased VLCFA levels, an effect that was dependent on the presence of PAS2 indicating that PTPLA activity repressed FA elongation. The two dehydratases have specific expression profiles in the root with PAS2, mostly restricted to the endodermis, while PTPLA was confined in the vascular tissue and pericycle cells. Comparative ectopic expression of PTPLA and PAS2 in their respective domains confirmed the existence of two independent elongase complexes based on PAS2 or PTPLA dehydratase that are functionally interacting.


Assuntos
Acetiltransferases/metabolismo , Arabidopsis/enzimologia , Acetiltransferases/genética , Arabidopsis/genética , Retículo Endoplasmático/enzimologia , Elongases de Ácidos Graxos , Mutação , Saccharomyces cerevisiae/genética
19.
Curr Biol ; 25(19): 2584-90, 2015 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-26387715

RESUMO

The phloem is a vascular strand that conducts photoassimilates and systemic signals throughout the plant to coordinate growth. To date, few molecular genetic determinants have been identified to control both specification and differentiation of this tissue [1-3]. Among them, OCTOPUS (OPS) protein was previously identified as a polarly localized plasma membrane-associated protein of unknown biochemical function whose broad provascular expression becomes restricted to the phloem upon differentiation [2]. OPS loss-of-function mutants showed an altered vascular network in cotyledons and an intermittent phloem differentiation in the root [2, 4]. Here, we demonstrate a role for OPS as a positive regulator of the brassinosteroid (BR) signaling pathway. Indeed, transgenic lines overexpressing OPS (OPS-OE) display the hallmarks of constitutively overactivated BR mutants. Physiological and genetic analyses place OPS as a positive regulator of the BR signaling pathway upstream of the key transcription factors BES1 and BZR1. Directed protein interactions with known BR signaling proteins identified BIN2, a GSK3 protein involved in multiple signaling pathways, as a partner of OPS. This interaction recruits BIN2 to the plasma membrane, thus preventing its inhibitory activity in the nucleus. Finally, both bikinin (a potent inhibitor of GSK3 [5]) treatment and downstream dominant mutants bes1-D [6] and bzr1-D [7] can rescue phloem defects of ops in the root. Together, our data show that OPS antagonizes BIN2 to promote phloem differentiation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de Membrana/metabolismo , Proteínas Quinases/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Brassinosteroides/metabolismo , Células Cultivadas , Cotilédone/metabolismo , Mutação , Floema/crescimento & desenvolvimento , Floema/metabolismo , Fosforilação , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas , Proteínas Quinases/genética , Transdução de Sinais
20.
Annu Rev Plant Biol ; 63: 381-407, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22224451

RESUMO

Plant cell walls have the remarkable property of combining extreme tensile strength with extensibility. The maintenance of such an exoskeleton creates nontrivial challenges for the plant cell: How can it control cell wall assembly and remodeling during growth while maintaining mechanical integrity? How can it deal with cell wall damage inflicted by herbivores, pathogens, or abiotic stresses? These processes likely require mechanisms to keep the cell informed about the status of the cell wall. In yeast, a cell wall integrity (CWI) signaling pathway has been described in great detail; in plants, the existence of CWI signaling has been demonstrated, but little is known about the signaling pathways involved. In this review, we first describe cell wall-related processes that may require or can be targets of CWI signaling and then discuss our current understanding of CWI signaling pathways and future prospects in this emerging field of plant biology.


Assuntos
Parede Celular/metabolismo , Células Vegetais/metabolismo , Desenvolvimento Vegetal/fisiologia , Proteínas de Arabidopsis/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Parede Celular/química , Glucanos/metabolismo , Pectinas/metabolismo , Fosfotransferases/metabolismo , Proteínas de Plantas/metabolismo , Transdução de Sinais/fisiologia , Água/metabolismo , Xilanos/metabolismo
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