Arabidopsis thaliana
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ABSTRACT:
PROVIDER: PRJNA145761 | ENA |
REPOSITORIES: ENA
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Project description:Pattern recognition receptors (PRRs) at the plasma membrane promote plant immunity through the detection of conserved microbe-associated molecular patterns (MAMPs). In plants, the PRR for bacterial flagellin (flg22) is encoded by the receptor kinase FLS2. One of the earliest MAMP responses is the rapid and transient increase of cytosolic calcium (Ca2+) ions, which is required for many of the well-described downstream responses, e.g. generation of reactive oxygen species (ROS) and the transcriptional activation of defence-associated genes. Despite its relevance, the molecular components regulating the Ca2+ burst remain largely unknown. Here, we show that the plasma membrane P2B-type Ca2+ ATPase ACA8 forms a dynamic complex with the PRR FLS2. ACA8 and its closest homologue ACA10 are required for immunity against virulent bacteria. Mutant aca8 aca10 plants are reduced in the flg22-induced Ca2+ burst, show reduced ROS production and exhibit altered transcriptional reprogramming. In particular, flg22-induced gene expression is elevated downstream of signalling mitogen-activated protein (MAP) kinases, but reduced downstream of the calcium-dependent protein (CDP) kinase cascade. These results demonstrate that the fine regulation of Ca2+ fluxes in the cytosol is critical for the coordination of the downstream MAMP responses and provide for the first time a link between the FLS2 receptor complex and signalling kinases via the secondary messenger Ca2+. ACA8 also interacted with the BRI1 and CLV1 receptor kinases, which correlated with the developmental phenotypes of aca8 aca10 mutants suggesting a broader role for Ca2+ ATPases in receptor-mediated signalling. We used Affymetrix Arabidopsis Tiling 1.0R Array to compare global transcript levels in 7 days-old sterile grown seedlings. Steady-state mRNA levels in total RNA samples of 7 days old sterile seedlings
2012-04-24 | E-GEOD-31517 | biostudies-arrayexpress
Project description:Pattern recognition receptors (PRRs) at the plasma membrane promote plant immunity through the detection of conserved microbe-associated molecular patterns (MAMPs). In plants, the PRR for bacterial flagellin (flg22) is encoded by the receptor kinase FLS2. One of the earliest MAMP responses is the rapid and transient increase of cytosolic calcium (Ca2+) ions, which is required for many of the well-described downstream responses, e.g. generation of reactive oxygen species (ROS) and the transcriptional activation of defence-associated genes. Despite its relevance, the molecular components regulating the Ca2+ burst remain largely unknown. Here, we show that the plasma membrane P2B-type Ca2+ ATPase ACA8 forms a dynamic complex with the PRR FLS2. ACA8 and its closest homologue ACA10 are required for immunity against virulent bacteria. Mutant aca8 aca10 plants are reduced in the flg22-induced Ca2+ burst, show reduced ROS production and exhibit altered transcriptional reprogramming. In particular, flg22-induced gene expression is elevated downstream of signalling mitogen-activated protein (MAP) kinases, but reduced downstream of the calcium-dependent protein (CDP) kinase cascade. These results demonstrate that the fine regulation of Ca2+ fluxes in the cytosol is critical for the coordination of the downstream MAMP responses and provide for the first time a link between the FLS2 receptor complex and signalling kinases via the secondary messenger Ca2+. ACA8 also interacted with the BRI1 and CLV1 receptor kinases, which correlated with the developmental phenotypes of aca8 aca10 mutants suggesting a broader role for Ca2+ ATPases in receptor-mediated signalling. We used Affymetrix Arabidopsis Tiling 1.0R Array to compare global transcript levels in 7 days-old sterile grown seedlings.
2012-04-25 | GSE31517 | GEO
Project description:Clathrin-mediated endocytosis is essential for a wide range of cellular functions. We used a multi- step siRNA-based screening strategy to identify novel regulators of the first step of clathrin- mediated endocytosis, formation of clathrin-coated vesicles (CCVs) at the plasma membrane. A primary genome-wide screen identified 334 hits that caused accumulation of CCV cargo on the cell surface. A secondary screen identified 92 hits that inhibited cargo uptake and/or altered the morphology of clathrin-coated structures. The hits include components of four functional complexes: coat proteins, V-ATPase subunits, spliceosome-associated proteins, and acetyltransferase subunits. Electron microscopy revealed that V-ATPase depletion caused the cell to form aberrant non-constricted clathrin-coated structures at the plasma membrane. The V- ATPase knockdown phenotype was rescued by addition of exogenous cholesterol, indicating that the knockdown blocks clathrin-mediated endocytosis by preventing cholesterol from recycling from endosomes back to the plasma membrane. The microarray analysis was performed to test whether the siRNA targets are expressed in the cell lines used in the screen. For the microarray analysis of gene expression, the two cell lines used in the study were analyzed in duplicate: HeLa-YXXΦ and HeLa-FXNPXY (Hela-M cells expressing a CD8-YXXΦ and CD8-FXNPXY constructs respectively). YXXΦ and FXNPXY are motifs for clathrin-mediated endocytosis.
2012-12-01 | E-GEOD-39954 | biostudies-arrayexpress
Project description:Clathrin-mediated endocytosis is essential for a wide range of cellular functions. We used a multi- step siRNA-based screening strategy to identify novel regulators of the first step of clathrin- mediated endocytosis, formation of clathrin-coated vesicles (CCVs) at the plasma membrane. A primary genome-wide screen identified 334 hits that caused accumulation of CCV cargo on the cell surface. A secondary screen identified 92 hits that inhibited cargo uptake and/or altered the morphology of clathrin-coated structures. The hits include components of four functional complexes: coat proteins, V-ATPase subunits, spliceosome-associated proteins, and acetyltransferase subunits. Electron microscopy revealed that V-ATPase depletion caused the cell to form aberrant non-constricted clathrin-coated structures at the plasma membrane. The V- ATPase knockdown phenotype was rescued by addition of exogenous cholesterol, indicating that the knockdown blocks clathrin-mediated endocytosis by preventing cholesterol from recycling from endosomes back to the plasma membrane. The microarray analysis was performed to test whether the siRNA targets are expressed in the cell lines used in the screen.
2012-12-01 | GSE39954 | GEO
Project description:Transcript profiling of Arabidopsis plasma membrane proton pump mutants
| PRJNA149843 | ENA
Project description:Plasma membrane (PM) H+-ATPase contributes to nutrient uptake and stomatal opening by creating proton gradient across the membrane. A dominant mutation in the OPEN STOMATA2 locus (OST2-2D) constitutively activates Arabidopsis PM H+-ATPase 1 (AHA1), enlarging proton motive force for root nutrient uptake. However, the stomatal opening is also constitutively enhanced in the ost2-2D, which results in drought hypersensitivity. Therefore, we postulated that the root-specific activation of OST2/AHA1 could be an ideal design for improving nutrient uptake efficiency without causing drought hypersensitivity. Accordingly, we grafted Col-0 (WT) scions onto rootstock originating from WT or ost2-2D and analyzed the vegetative growth, nutrient element content, and transcriptomes of the grafted plants grown under nutrient-rich or -poor conditions.
2022-06-22 | E-MTAB-11843 | biostudies-arrayexpress
Project description:A highly conserved bacterial effector localizes to the host plasma membrane and downregulates the expression of the NDR/HIN1-like 13 (NHL13) gene requiried for antibacterial immunity in Arabidopsis
| PRJNA279184 | ENA
Project description:Effect of flg22 treatment in Arabidopsis thaliana Ecotype Landsberg versus fls2-17 mutant
2004-08-01 | E-MEXP-81 | biostudies-arrayexpress
Project description:To compare transcriptional outputs of FLS2 and SGN3 pathways solely within the endodermis, we conducted comparative RNAseq analysis of pCASP1::FLS2 fls2 and wild-type seedling roots treated by flg22 and CIF2 respectively, at three timepoints (30, 120, 480 min) (CIF2 responses data previously published by Fujita et al 2020). The receptor mutant controls sgn3 and fls2, displayed minimal gene responses to respective peptides.
2024-08-11 | GSE270950 | GEO
Project description:In this study, we conducted protoplast-based gain-of-function assay to screen 23 members of the RLCK VII family for those whose overexpression can affect flg22-induced MAPK activation. We identified CDG1, a previously characterized positive regulator in brassinosteroid (BR) signaling (Kim et al., 2011), as a new negative regulator of flg22-triggered MAPK activation. We further found that CDG1 could constitutively associate with FLS2 and CERK1, and its transgenic overexpression could significantly reduce the abundances of these two essential PRRs, leading to impaired immune responses to both flg22 and chitin as well as increased susceptibility to bacterial and fungal pathogens. Surprisingly, the CDG1-mediated degradation of FLS2 and CERK1 is independent of E3 ubiquitin ligases PUB12/PUB13, but requires the kinase activity of MEKK1, a MAPKKK positively regulating plant innate immunity (Asai et al., 2002). Moreover, we demonstrated that RIN4 and the P. syringae effector AvrRpm1 could also interact with CDG1, and obtained evidence that CDG1 may be one of the RLCK VII members responsible for the AvrRpm1-induced RIN4 phosphorylation. Our data thus reveal multifaceted moonlight function of CDG1 in Arabidopsis innate immunity in addition to its originally characterized role in BR signaling.
2019-05-22 | GSE130958 | GEO