A cell type-specific proteomics uncovers a guard cell-specific RAF15-SnRK2.6/OST1 kinase cascade
Ontology highlight
ABSTRACT:
ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)
SUBMITTER: Pengcheng Wang
PROVIDER: PXD034026 | JPOST Repository | Tue May 23 00:00:00 BST 2023
REPOSITORIES: jPOST
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| 20200717_whl_GCP1.msf | Msf | |||
| 20200717_whl_GCP1.raw | Raw | |||
| 20200717_whl_GCP1.xlsx | Xlsx | |||
| 20200717_whl_GCP2.msf | Msf | |||
| 20200717_whl_GCP2.raw | Raw |
Items per page: 5 1 - 5 of 18 |
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Project description:Multicellular organisms such as plants contain different types of cells with specialized functions. Analyzing the characteristics of each type of cells will reveal specific cell functions and enhance understanding of how an organism organizes and works. Here we report a high-sensitive and efficient cell-type-specific multi-omics pipeline, combining simplified flow cytometry-based fluorescent cell-sorting for fluorescent protoplasts and optimized nano-scale proteomics and metabolomics methods, which allow us to in-depth analyze the proteomics and metabolomics of a particular type of cells. By this method, we quantitatively compared the proteomics and metabolomics between guard cells and mesophyll cells and revealed that the enrichment of signal transduction-related proteins enables guard cells to respond to various environmental stimuli quickly. We uncovered a guard-cell-specific kinase cascade consisting of RAF15 and OST1 mediates the ABA-induced stomatal closure in guard cells. This pipeline is applicable to various types of cells in plant or non-plant systems to acquire systemic knowledge of how cells work specifically and in highly organized multiple cell organisms.
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Project description:Multicellular organisms such as plants contain different types of cells with specialized functions. Analyzing the characteristics of each type of cells will reveal specific cell functions and enhance understanding of how an organism organizes and works. Here we report a high-sensitive and efficient cell-type-specific multi-omics pipeline, combining simplified flow cytometry-based fluorescent cell-sorting for fluorescent protoplasts and optimized nano-scale proteomics and metabolomics methods, which allow us to in-depth analyze the proteomics and metabolomics of a particular type of cells. By this method, we quantitatively compared the proteomics and metabolomics between guard cells and mesophyll cells and revealed that the enrichment of signal transduction-related proteins enables guard cells to respond to various environmental stimuli quickly. We uncovered a guard-cell-specific kinase cascade consisting of RAF15 and OST1 mediates the ABA-induced stomatal closure in guard cells. This pipeline is applicable to various types of cells in plant or non-plant systems to acquire systemic knowledge of how cells work specifically and in highly organized multiple cell organisms.
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Project description:Multicellular organisms such as plants contain different cell types with specialized functions. Analyzing the characteristics of each cell type reveals specific cell functions and enhances understanding of organization and function at the organismal level. Guard cells are specialized epidermal cells that play an important role in gaseous exchange in and out of plant leaves by regulating the opening and closing of the stomata, and provide a model genetic system for analyzing cell fate, signaling and function. Several proteomics analyses of guard cells are available, but these are limited in depth. Here we used enzymatic isolation and flow cytometry to enrich for guard cell and mesophyll cell protoplasts and acquire in-depth proteomics in these two major cell types in Arabidopsis leaves. We identified ~3,000 proteins not previously found in the guard cell proteome and more than 500 proteins that may be specific to guard cells. The depth of our proteomics enabled us to uncover a guard cell-specific kinase cascade whereby Raf-like kinase15(RAF15) and Snf1-related kinase2.6(SnRK2.6)/OST1(open stomata 1) mediate abscisic acid (ABA)-induced stomatal closure. RAF15 directly phosphorylated SnRK2.6/OST1 at the conserved Ser175 residue in its activation loop and was sufficient to reactivate the dephosphorylated inactive form of SnRK2.6/OST1. ABA-triggered SnRK2.6/OST1 activation and stomatal closure was impaired in raf15 mutants. Our study answers the long-standing question of how ABA activates SnRK2.6/OST1 in guard cells and represents a resource potentially providing many more insights into the molecular basis of guard cell and mesophyll cell development, structure, and function.
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Project description:Multicellular organisms such as plants contain different cell types with specialized functions. Analyzing the characteristics of each cell type reveals specific cell functions and enhances understanding of organization and function at the organismal level. Guard cells are specialized epidermal cells that play an important role in gaseous exchange in and out of plant leaves by regulating the opening and closing of the stomata, and provide a model genetic system for analyzing cell fate, signaling and function. Several proteomics analyses of guard cells are available, but these are limited in depth. Here we used enzymatic isolation and flow cytometry to enrich for guard cell and mesophyll cell protoplasts and acquire in-depth proteomics in these two major cell types in Arabidopsis leaves. We identified ~3,000 proteins not previously found in the guard cell proteome and more than 500 proteins that may be specific to guard cells. The depth of our proteomics enabled us to uncover a guard cell-specific kinase cascade whereby Raf-like kinase15(RAF15) and Snf1-related kinase2.6(SnRK2.6)/OST1(open stomata 1) mediate abscisic acid (ABA)-induced stomatal closure. RAF15 directly phosphorylated SnRK2.6/OST1 at the conserved Ser175 residue in its activation loop and was sufficient to reactivate the dephosphorylated inactive form of SnRK2.6/OST1. ABA-triggered SnRK2.6/OST1 activation and stomatal closure was impaired in raf15 mutants. Our study answers the long-standing question of how ABA activates SnRK2.6/OST1 in guard cells and represents a resource potentially providing many more insights into the molecular basis of guard cell and mesophyll cell development, structure, and function.
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Project description:Multicellular organisms such as plants contain different cell types with specialized functions. Analyzing the characteristics of each cell type reveals specific cell functions and enhances understanding of organization and function at the organismal level. Guard cells are specialized epidermal cells that play an important role in gaseous exchange in and out of plant leaves by regulating the opening and closing of the stomata, and provide a model genetic system for analyzing cell fate, signaling and function. Several proteomics analyses of guard cells are available, but these are limited in depth. Here we used enzymatic isolation and flow cytometry to enrich for guard cell and mesophyll cell protoplasts and acquire in-depth proteomics in these two major cell types in Arabidopsis leaves. We identified ~3,000 proteins not previously found in the guard cell proteome and more than 600 proteins that may be specific to guard cells. The depth of our proteomics enabled us to uncover a guard cell-specific kinase cascade whereby Raf-like kinase15(RAF15) and Snf1-related kinase2.6(SnRK2.6)/OST1(open stomata 1) mediate abscisic acid (ABA)-induced stomatal closure. RAF15 directly phosphorylated SnRK2.6/OST1 at the conserved Ser175 residue in its activation loop and was sufficient to reactivate the dephosphorylated inactive form of SnRK2.6/OST1. ABA-triggered SnRK2.6/OST1 activation and stomatal closure was impaired in raf15 mutants. We also showed that enrichment of enzymes and in flavone metabolism in GCP, and consistently, dramatically highly accumulation of flavone metabolites. Our study answers the long-standing question of how ABA activates SnRK2.6/OST1 in guard cells and represents a resource potentially providing many more insights into the molecular basis of guard cell and mesophyll cell development, metabolism, structure, and function.
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