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Arabidopsis thaliana gene expression in adult leaf tissue of mutants in exocyst subunits exo70B1, exo70B2 and exo70B1xexo70B2

ABSTRACT: Several mutants in plant exocyst vesicle-tethering complex have various phenotypes. We decided to look into the differential gene expression for subunits exo70B1, B2 and double B1xB2 mutant, which were found before to be mores susceptible toward Pseudomonas in young developmental stages, but may develop enhanced resistance in later stages of development (especially exo70B1). We uncovered both the quantitative and qualitative changes in gene expressions among the analysed mutant lines in comparison to WT. We discovered several interesting candidate genes involved in stomatal and cell wall function, that would be worth of further more focused studies.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE311031 | GEO | 2025/11/30

REPOSITORIES: GEO

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Arabidopsis thaliana gene expression in seedlings of mutants in exocyst subunits exo70B1, exo70B2 and exo70B1xexo70B2

Project description:Several mutants in plant exocyst vesicle-tethering complex have various phenotypes. We decided to look into the differential gene expression for subunits exo70B1, B2 and double B1xB2 mutant, which were found before to be mores susceptible toward Pseudomonas in young developmental stages. We uncovered both the quantitative and qualitative changes in gene expressions among the analysed mutant lines in comparison to WT. We discovered several interesting candidate genes involved in cell wall function, that would be worth of further more focused studies.

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Mutations in the exocyst component EXOC2 (Sec5) cause severe defects in human brain development

Project description:The exocyst is an octameric protein complex and an essential component of the membrane transport machinery required for tethering vesicles at the plasma membrane prior to fusion. Here we report on pathogenic variants in one of the exocyst subunits, EXOC2 (Sec5), in unrelated families. Analysis of patient’s fibroblasts revealed reduced protein levels in Family 1, impaired secretory vesicle fusion with the plasma membrane and defective bulk exocytosis. Based on genetic and functional genomics findings, we suggest that the EXOC2 variants we have identified are the cause of the neurological disorder in these patients.

2020-06-29 | PXD017107 | Pride

Exocyst Dynamics During Vesicle Tethering and Fusion

Project description:The exocyst is a conserved octameric complex that tethers exocytic vesicles to the plasma membrane prior to fusion. Exocyst assembly and delivery mechanisms remain unclear, especially in mammalian cells. Here we tagged multiple endogenous exocyst subunits with sfGFP or Halo using Cas9 gene-editing, to create single and double knock-in lines of mammary epithelial cells, and interrogated exocyst dynamics by high-speed imaging and correlation spectroscopy. We discovered that mammalian exocyst is comprised of tetrameric subcomplexes that can associate independently with vesicles and plasma membrane and are in dynamic equilibrium with octamer and monomers. Membrane arrival times are similar for subunits and vesicles, but with a small delay (~80msec) between subcomplexes. Departure of SEC3 occurs prior to fusion, whereas other subunits depart just after fusion. About 9 exocyst complexes are associated per vesicle. These data reveal the mammalian exocyst as a remarkably dynamic two-part complex and provide important insights into assembly/disassembly mechanisms.

2018-12-05 | PXD011370 | Pride

Disruption of the exocyst induces podocyte loss and dysfunction

Project description:Disruption of the exocyst induces podocyte loss and dysfunction

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Pancreatic Beta-Cell Enhancers Regulate Rhythmic Transcription of Exocyst Triggering and Diabetes [ChIP-seq]

Project description:The molecular clock is a transcriptional oscillator present in brain and peripheral cells that coordinates behavior and physiology with the solar cycle. Here we reveal that the clock gates insulin secretion through genome-wide transcriptional control of the pancreatic exocyst across species. Clock transcription factors bind to unique enhancer sites in cycling genes in beta cells that diverge from those in liver, revealing the dynamics of inter-tissue clock control of genomic and physiologic processes important in glucose homeostasis.

2015-11-10 | GSE70960 | GEO

Pancreatic Beta-Cell Enhancers Regulate Rhythmic Transcription of Exocyst Triggering and Diabetes [ChIP-seq]

2015-11-10 | E-GEOD-70960 | biostudies-arrayexpress

Fission yeast NDR/LATS kinase Orb6 regulates exocytosis via phosphorylation of exocyst complex

Project description:NDR/LATS kinases regulate multiple aspects of cell polarity and morphogenesis from yeast to mammals, but few of their substrates are known. Fission yeast NDR/LATS kinase Orb6 has been proposed to control cell polarity via spatial regulation of Gef1, a guanine nucleotide exchange factor for the small GTPase Cdc42. Here we show that Orb6 plays a critical role as a positive regulator of exocytosis, independent of Gef1. Through Orb6 inhibition in vivo and quantitative global phosphoproteomics, we identify several proteins involved in membrane trafficking as Orb6 targets, and we confirm Sec3 and Sec5, conserved components of the exocyst complex, as substrates of Orb6 both in vivo and in vitro. Our results suggest that Orb6 kinase activity is crucial for exocyst localization to actively-growing cell tips and for exocyst activity during septum dissolution after cytokinesis. We further show that Orb6 phosphorylation of Sec3 serine-201 contributes to exocyst function in parallel with exocyst protein Exo70. We propose that Orb6 contributes to polarized growth by regulating membrane trafficking at multiple levels.

2019-01-31 | PXD009408 | Pride

Conservation of helical bundle structure between the exocyst subunits.

Project description:BackgroundThe exocyst is a large hetero-octomeric protein complex required for regulating the targeting and fusion of secretory vesicles to the plasma membrane in eukaryotic cells. Although the sequence identity between the eight different exocyst subunits is less than 10%, structures of domains of four of the subunits revealed a similar helical bundle topology. Characterization of several of these subunits has been hindered by lack of soluble protein for biochemical and structural studies.Methodology/principal findingsUsing advanced hidden Markov models combined with secondary structure predictions, we detect significant sequence similarity between each of the exocyst subunits, indicating that they all contain helical bundle structures. We corroborate these remote homology predictions by identifying and purifying a predicted domain of yeast Sec10p, a previously insoluble exocyst subunit. This domain is soluble and folded with approximately 60% alpha-helicity, in agreement with our predictions, and capable of interacting with several known Sec10p binding partners.Conclusions/significanceAlthough all eight of the exocyst subunits had been suggested to be composed of similar helical bundles, this has now been validated by our hidden Markov model structure predictions. In addition, these predictions identified protein domains within the exocyst subunits, resulting in creation and characterization of a soluble, folded domain of Sec10p.

| S-EPMC2635961 | biostudies-literature

Exocyst Subunit Exo70B2 is Linked to Immune Signalling and Autophagy

Project description:During the immune response, activation of the secretory pathway is key to mount an effective response, while gaging its output is important to maintain cellular homeostasis. Exo70 subunit of the exocyst functions as a spatiotemporal regulator by mediating numerous interactions with proteins and lipids. However, a molecular understanding of the exocyst regulation remains challenging. We show that the Exo70B2 behaves as a bona fide exocyst subunit. Conversely, treatment with the salicylic acid (SA) defence hormone analogue Benzothiadiazole (BTH), or the immunogenic peptide flg22, induced Exo70B2 transport into the vacuole. We reveal that Exo70B2 interacts with ATG8 via two ATG8-interacting motives (AIMs) and its transport into the vacuole was dependent on autophagy. In line with its role in immunity, we discovered that Exo70B2 interacted with and was phosphorylated by the kinase MPK3. Mimicking phosphorylation had a dual impact on Exo70B2; first, by inhibiting localization at sites of active secretion, and second, it increased the interaction with ATG8. Phosphonull variants displayed higher Effector-Triggered Immunity (ETI) and were hypersensitive to BTH, which induce secretion and autophagy. Our results suggest a molecular mechanism by which phosphorylation diverts Exo70B2 from the secretory into the autophagy pathway for its degradation, to dampen secretory activity.

2021-03-01 | PXD022246 | Pride

Microarray analysis of scap1 mutant and wild type in Arabidopsis thaliana guard cell protoplast

Project description:Stomata are highly specialized organs which consist of pairs of guard cells and regulate gas and water vapor exchange in plants. While early stages of guard cell differentiation have been described and were interpreted in analogy to processes of cell type differentiation in animals, the downstream development of functional stomatal guard cells remains poorly understood. We have isolated an Arabidopsis mutant, scap1 (stomatal carpenter 1), that develops irregularly shaped guard cells and lacks the ability to control stomatal aperture, including CO2-induced stomatal closing and light-induced stomatal opening. SCAP1 was identified as a plant-specific Dof-type transcription factor expressed in maturing guard cells but not in guard mother cells. SCAP1 regulates the expression of genes encoding key elements of stomatal functioning and morphogenesis, such as a K+ channel protein, MYB60 transcription factor, and pectin methyl esterase. Consequently, ion homeostasis was disturbed in scap1 guard cells, and esterification of extracellular pectins was impaired so that the cell walls lining the pores did not mature normally. We conclude that SCAP1 regulates essential processes of stomatal guard cell maturation and functions as a key transcription factor regulating the final stages of guard cell differentiation.

2013-02-28 | GSE43964 | GEO

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