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Effectors of RAB GTPases and their role in plant secretion
Růžičková, Martina ; Hála, Michal (advisor) ; Motyka, Václav (referee) ; Klíma, Petr (referee)
Rab GTPases are small signaling molecules that play an important role in vesicle trafficking in eukaryotic cells. Correct signaling through small GTPases allows orchestration of vesicle transport among cellular organelles and also to the cell wall providing cell wall material for cell growth and elongation. Engagement of Rab GTPases in the regulation of endomembrane trafficking is one of the evolutionary conserved aspects of secretion regulation. The network of Rab GTPases interaction includes also various downstream effectors. One of them is the exocyst complex involved in vesicle docking at the plasma membrane. It is a complex composed of eight different subunits (Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70 and Exo84). Exocyst was discovered as Sec4p Rab GTPase effector in yeast and also data from animal models describe the Sec15 exocyst subunit as the Rab-interacting partner, but data from plants are missing. On the other hand, numerous studies identified exocyst role in tip growth of pollen tube and root hairs, seed coat formation, cell plate and cell wall formation, hypocotyl elongation, and importantly also PIN auxin efflux carriers recycling and polar auxin transport. There are two paralogues of SEC15 in the Arabidopsis genome, SEC15a and SEC15b, the previous one already shown to be...
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Functions of RAB GTPases and SNARE proteins in post-Golgi trafficking pathways in response to abiotic stress in plants
Moulík, Michal ; Hála, Michal (advisor) ; Mašková, Petra (referee)
Plants as sessile organisms are strongly affected by abiotic and biotic stress factors. Thus, they have developed an array of morfological, biochemical and physiological adaptations to reduce the negative effects of these factors. The membrane trafficking, among others, plays very important role in adaptation to abiotic stress. In my bachelor thesis I have focused on two important protein families involved in this trafficking, namely on RAB GTPases and SNARE proteins. In the first part, the phenomenon of stress is characterized and the strategies how plants cope with the effect of stressors are described, especially the production of reactive oxygen species (ROS) and autophagy. Following chapter deals with an introduction to the membrane trafficking in plants. In the rest of the thesis, I characterize RAB GTPases and SNARE proteins and provide contemporary insight in the mechanism of their function. The aim of the key parts of these chapters is to summarize current knowledge of RAB GTPases' and SNARE proteins' functions in post-Golgi trafficking pathways during response to abiotic stressors or secondary oxidative stress. Key words: plants, abiotic stress, membrane trafficking, secretion, endocytosis, vacuole, RAB GTPases, SNARE proteins
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Effectors of RAB GTPases and their role in plant secretion
Růžičková, Martina ; Hála, Michal (advisor) ; Motyka, Václav (referee) ; Klíma, Petr (referee)
Rab GTPases are small signaling molecules that play an important role in vesicle trafficking in eukaryotic cells. Correct signaling through small GTPases allows orchestration of vesicle transport among cellular organelles and also to the cell wall providing cell wall material for cell growth and elongation. Engagement of Rab GTPases in the regulation of endomembrane trafficking is one of the evolutionary conserved aspects of secretion regulation. The network of Rab GTPases interaction includes also various downstream effectors. One of them is the exocyst complex involved in vesicle docking at the plasma membrane. It is a complex composed of eight different subunits (Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70 and Exo84). Exocyst was discovered as Sec4p Rab GTPase effector in yeast and also data from animal models describe the Sec15 exocyst subunit as the Rab-interacting partner, but data from plants are missing. On the other hand, numerous studies identified exocyst role in tip growth of pollen tube and root hairs, seed coat formation, cell plate and cell wall formation, hypocotyl elongation, and importantly also PIN auxin efflux carriers recycling and polar auxin transport. There are two paralogues of SEC15 in the Arabidopsis genome, SEC15a and SEC15b, the previous one already shown to be...
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Endocytic transport in cytokinesis
Koudelová, Kristina ; Libusová, Lenka (advisor) ; Vosolsobě, Stanislav (referee)
Cytokinesis represents a very complex and highly orchestrated process. For many years, the mechanism of animal cell cytokinesis was described as a result of actomyosin ring constriction. By contrast, in plant cells the division was seen as an outcome of vesicle fusion at the cell plate region between two daughter cells. Recent studies, however, uncover the involvement of vesicular trafficking in animal cell cytokinesis. This thesis aims to highlight the importance of endocytic transport and the necessity of its proper regulation. At first, the origin of vesicles is debated. Afterwards, three main types of endocytic vesicles are examined - Rab11/FIP3 endosomes, Rab35-endosomes and PI(3)P-enriched endosomes, along with their function and interacting partners. Finally, the attention is given to the mechanism of abscission and midbody inheritance. Ongoing processes are accompanied by changes in membrane composition, cytoskeleton reorganization and targeted delivery of distinct cargo molecules. Failure in cytokinesis has been implicated in the etiology of many diseases, such as cancer. Therefore, better understanding of associated endocytic trafficking may provide us with new therapeutic strategies.
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The role of cytoskeleton in endosomal fusion and fission
Získalová, Tereza ; Libusová, Lenka (advisor) ; Tolde, Ondřej (referee)
Cytoskeleton plays a key role in endocytic process. Vesicules move along microtubules to target membranes. Microtubules also partake in the formation of endosomal tubules, from which recyclated vesicules are splitted off. Actin network has in endocytosis multi-ple effect as well. In the case of membrane fusion is its role both, positive and negative, for it creates mechanical force which facilitates the fusion in last stage. By contrast, in the first stage, it acts as a physical barrier, which needs to be removed. Actin also actively participates in fission of vesicules. Actin network and microtubules are thus interconnected with endocytic pathway in time and space. Right functional connection of the cytoskeleton with dynamics of endocytic vesicles is driven by many regulatory proteins. Among important regulators of actin network belong for example proteins of Arp2/3, WASH complex, WASP or Rab and Rho proteins. Powered by TCPDF (www.tcpdf.org)
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Characterization of selected RAB GTPase activating protein (RAB GAP) of Arabidopsis thaliana
Metlička, Jáchym ; Hála, Michal (advisor) ; Eliáš, Marek (referee)
8 ABSTRACT Rab GTPases (Rabs) are the most populous branch of eukaryotic Ras GTPase superfamily. In active GTP-binding conformation, they serve as key instruments in defining transient membrane identity and through various effectors regulate formation, transport, conversion, and fusion of membrane vesicles. This is important for upkeep of compartmentalized structure of eukaryotic cells and for facilitating both endo- and exocytic processes. Rabs are converted into GDP-binding conformation by interactions with Rab GTPase activating proteins (Rab GAPs) that possess ability to significantly speed up weak intrinsic GTP hydrolytic activity of Rabs. Through this process, Rab GAPs can limit scope of the Rabs' activity and lay out spatiotemporal boundaries for varying Rab populations. In this thesis, I tried to characterize a Rab GAP, GAP2, seemingly necessary for standard development of thale cress plants. Besides TBC catalytic domain, GAP2 (product of At2g39280 gene) possesses a C-terminal coiled-coil motif, which was previously found to interact with Rab GTPases. Experiments aiming to complement T-DNA insertion mutant in GAP2, elucidate GAP2 intracellular localization, novel interacting partners, and character of interaction with the Rabs discovered in the pilot study were undertaken. The results suggest that...
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