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Nuclear import and export inhibitors analysis in plants
Haltuf, Josef ; Schwarzerová, Kateřina (advisor) ; Angelini, Jindřiška (referee)
Eukaryotic cells have a nucleus where DNA is stored. Transport between the nucleus and cytoplasm is extremely important for ensuring gene expression as well as for controlling and regulating nuclear processes and structures. During interphase, thousands of macromolecules are transported into and out of the nucleus through nuclear pores, and nucleocytoplasmic transport is essential for the viability of eukaryotic organisms. Transport between the nucleus and cytoplasm is tightly regulated, and specific inhibitors of nuclear transport are used to study this regulation. While nuclear transport inhibitors have been extensively studied in animal cell research, the activity of most of them has not been tested in plant cells. In this work the nuclear export inhibitors Leptomycin B (LMB) and the nuclear import inhibitors Importazole (IMZ) and Ivermectin (IVR) are tested in Arabidopsis thaliana plant roots and in isolated tobacco BY- 2 cells. This work also includes testing suitable markers for transport between the nucleus and cytoplasm. GFP-NES and NLS-GFP were found to be the best markers because they are stable and can diffuse spontaneously through nuclear pores. With these markers, I determined effective concentrations for LMB (100 nM in roots, 50 nM in isolated cells) and IMZ (75 μM in roots). IVR proved to be...
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The Role of Cytoskeleton and Phospholipids in signaling During Plant Defense Response
Angelini, Jindřiška ; Dušková, Jiřina (advisor) ; Jahodář, Luděk (referee) ; Grančai, Daniel (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Department of pharmaceutical botany and ecology Candidate PharmDr. Jindřiška Matoušková Supervisor Ass. Prof. RNDr. Jiřina Dušková, Csc Title of Doctoral Thesis The role of cytoskeleton and phospholipids in signaling during plant defense response Upon abiotic and biotic cues, plant surveillance system simultaneously triggers a variety of signaling events to develop an effective defense response within a few hours. On the molecular level, signaling period is the chain of many consecutive and fast biochemical reactions. Tracking of one particular signaling event e.g. during plant-pathogen interactions is quite difficult task. Presented work is focused on several components of this intricate defense network; namely on the role of salicylic acid, phospholipid signaling and cytoskeleton dynamics and their mutual interaction coupled with downstream changes of plant metabolom. Live imaging can help to elucidate localization and dynamics of particular, fluorescently labeled proteins during defense response. Plenty of important events occurring in plant- -pathogen interactions take place in plasma membrane: where pathogen perception, exocytosis, endocytosis occure and ion channels and carriers are located. Also phosphatidic...
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The Role of Cytoskeleton and Phospholipids in signaling During Plant Defense Response
Angelini, Jindřiška ; Dušková, Jiřina (advisor) ; Jahodář, Luděk (referee) ; Grančai, Daniel (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Department of pharmaceutical botany and ecology Candidate PharmDr. Jindřiška Matoušková Supervisor Ass. Prof. RNDr. Jiřina Dušková, Csc Title of Doctoral Thesis The role of cytoskeleton and phospholipids in signaling during plant defense response Upon abiotic and biotic cues, plant surveillance system simultaneously triggers a variety of signaling events to develop an effective defense response within a few hours. On the molecular level, signaling period is the chain of many consecutive and fast biochemical reactions. Tracking of one particular signaling event e.g. during plant-pathogen interactions is quite difficult task. Presented work is focused on several components of this intricate defense network; namely on the role of salicylic acid, phospholipid signaling and cytoskeleton dynamics and their mutual interaction coupled with downstream changes of plant metabolom. Live imaging can help to elucidate localization and dynamics of particular, fluorescently labeled proteins during defense response. Plenty of important events occurring in plant- -pathogen interactions take place in plasma membrane: where pathogen perception, exocytosis, endocytosis occure and ion channels and carriers are located. Also phosphatidic...
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