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Role of TCTP1 in plant reproduction
Pitoňak, Oliver ; Hafidh, Said (advisor) ; Lafon Placette, Clément (referee)
TCTP is a conserved eukaryotic protein involved in regulation of multiple cellular processes, such as translation, cell cycle and cell death. Apart from its intracellular functions, TCTP is secreted and participates in human immune response. Dimerization via a terminal cysteine residue is critical for its extracellular function. Multiple sequence alignment of eukaryotic TCTPs revealed that terminal cysteine residue is conserved not only in animals but also in green plant lineage. In contrast to animal model organisms, knowledge about TCTP in plants is limited. Arabidopsis thaliana genome harbors two TCTP paralogs, TCTP1 and TCTP2. TCTP1 expression is highly upregulated in pollen. Previously published studies pointed to its role in embryo development, pollen tube growth and targeting. Using mutant characterization and analysis of subcellular localization, the role of TCTP1 in plant reproduction was investigated in this thesis. Recombinant Arabidopsis thaliana TCTP1 protein was produced along with its potential interacting partners BRL2 and TTL3 in heterologous Escherichia coli system. tctp-1 T- DNA line characterization pointed to TCTP1 role in pollen tube growth. It was expressed in mature pollen and pollen tube under native promoter and localized to the cytoplasm. Recombinant AtTCTP1 formed dimers...
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It takes two to tango; signal perception during male-female interaction in flowering plants
Vlasák, Jonáš ; Honys, David (advisor) ; Vaňková, Radomíra (referee)
This thesis summarizes the current understanding of the communication between partners during the progamic phase of pollen with focus on the cruciferous family After attachment of conspecific pollen to stigma, pollen coat peptides (PCP) competitively inhibit the signaling perceived by the FERONIA receptor in the papillary cell and thus allow pollen acceptance. Contrary, the interaction of pollen protein SCR (S-LOCUS CYSTEIN RICH PROTEIN) with SRK receptor (S-LOCUS RECEPTOR KINASE) determines pollen incompatibility. The regulation of oriented secretion is essential for pollen tube (PT) growth. During the growth of the PT, the components of the extracellular matrix as well as stiffness and preassure are perceived. On its way to the egg, the PT matures and acquires the competence to perceive signals. The PT exits the transmitting tract thanks to the ovular signalisation. The PT is accepted by synergids, communication takes place involving RALF peptides (RAPID ALKALIZATION FACTOR) and the FERONIA receptor. The signaling results in the rupture of the PT and the transport of sperm cells to the female gametes. Sperm cells bind to the egg cell and the central cell and after confirmation, fuse with the female gametes. After fertilization, the egg cell degrades signaling molecules and the persistent synergid also...
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Function of phospholipases D and lipid phosphate phosphatases in the regulation of plant cell morphogenesis
Bezvoda, Radek ; Žárský, Viktor (advisor) ; Hašek, Jiří (referee) ; Vaňková, Radomíra (referee)
of the thesis The presented work explores the function and regulation of intracellular signaling that utilizes phospholipase D (PLD) and phosphatidic acid (PA), especially in the context of cellular morphogenesis of plants. PLDs cleave membrane phospholipids to phosphatidic acid, which has important biophysical and signaling role in many contexts, such as stress response, regulation of cytoskeletal dynamics and vesicular transport. Vesicular transport is essential in focused tip growth of plant pollen tubes and root hairs. Part of the work deals with NADPH oxidases, that are an emerging counterpart of PLD/PA signaling. Tobacco pollen tubes served as the main experimental model, as it enables assessing of changes in secretory pathway after pharmacological or genetic treatments. A technique utilizing antisense oligonucleotides was used for selective knock-down of PLD isoforms, NADPH oxidase and newly studied family of lipid phosphate phosphatases (LPPs) in pollen tubes. This enabled to assess functions of individual isoforms. For studying of selected gene families, various bioinformatic tool were utilized, such as dendrogram construction, analysis of available expression data and creating of virtual proteome. These tools together enabled to select potentially important genes for further experimental...
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Characterization of eukaryotic translation initiation factor 3 subunits (eIF3) in A. thaliana male gametophyte
Linhart, Filip ; Hafidh, Said (advisor) ; Retzer, Katarzyna (referee)
From RNA-to-protein, translation initiation and protein synthesis is mediated by trans-acting factors that recognize mRNA features common to almost all eukaryotes. Eukaryotic translation initiation factor 3 complex (eIF3) is a highly conserved protein complex that recognizes 5'-CAP elements of the mRNA to initiate translation. eIF3 consists of nine subunits, three of them having two isoforms: eIF3A, eIF2B1, eIF3B2, eIF3C1, eIF3C2, eIF3D, eIF3E, eIF3F, eIF3G1, eIF3G2, eIF3H and eIF3K. This work deals with functional characterization, expression and subcellular localization of eIF3B1, eIF3B2 and eIF3E in Arabidopsis thaliana male gametophyte and interaction of eIF3E with the Constitutive photomorphogenesis 9 (COP9) complex as a regulatory complex of eIF3E post-translational control. Here we show that depletion of eif3b1 or eif3b2 is not gametophytic lethal and that the two protein might function redundantly, whereas, knockout of eIF3E causes male gametophyte lethality. Interestingly, eif3b1 show post-fertilization defects during embryogenesis, suggesting that its redundancy with eIF3B2 is restricted to the gametophyte. Gene expression studies revealed high expression of eIF3 subunits in actively dividing zones of leaf primordia, root meristem and root elongation zones as well as in the vegetative...
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Molecular mechanisms of cell polarity and morphogenesis in moss Physcomitrella patens
D'Agostino, Viktoria ; Žárský, Viktor (advisor) ; Soukup, Aleš (referee)
Plant cells are able to establish polarity and expand by tip growth. Polarized cells often embrace functions important for plant viability. The process of tip growth requires actin cytoskeleton in collaboration with a number of accessory proteins. The position of the intensively expanding region is provided by microtubules and the function of signalling proteins. Polarized secretion regulates the structural properties and subsequently the shape of the cell wall. Some components of the secretory and signalling pathways are highly conserved among eukaryotes, others are found exclusively in the plant kingdom. Though much has been discovered in yeast and animal cells, many mechanisms in plants are yet to be revealed. Model systems performing tip growth, such as root hairs, pollen tubes and protonema cells, enable comparison and thus a complementary overview of the various processes.
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Function of phospholipases D and lipid phosphate phosphatases in the regulation of plant cell morphogenesis
Bezvoda, Radek ; Žárský, Viktor (advisor) ; Hašek, Jiří (referee) ; Vaňková, Radomíra (referee)
of the thesis The presented work explores the function and regulation of intracellular signaling that utilizes phospholipase D (PLD) and phosphatidic acid (PA), especially in the context of cellular morphogenesis of plants. PLDs cleave membrane phospholipids to phosphatidic acid, which has important biophysical and signaling role in many contexts, such as stress response, regulation of cytoskeletal dynamics and vesicular transport. Vesicular transport is essential in focused tip growth of plant pollen tubes and root hairs. Part of the work deals with NADPH oxidases, that are an emerging counterpart of PLD/PA signaling. Tobacco pollen tubes served as the main experimental model, as it enables assessing of changes in secretory pathway after pharmacological or genetic treatments. A technique utilizing antisense oligonucleotides was used for selective knock-down of PLD isoforms, NADPH oxidase and newly studied family of lipid phosphate phosphatases (LPPs) in pollen tubes. This enabled to assess functions of individual isoforms. For studying of selected gene families, various bioinformatic tool were utilized, such as dendrogram construction, analysis of available expression data and creating of virtual proteome. These tools together enabled to select potentially important genes for further experimental...
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Interplay of cytoskeleton and secretory pathway during exocytosis in plant cells
Aldorfová, Klára ; Sekereš, Juraj (advisor) ; Vosolsobě, Stanislav (referee)
Cytoskeleton is known to participate in exocytosis of yeast and animal cells. The role of plant cytoskeleton during exocytosis has not been fully understood yet. However, both actin and microtubules evidently contributes to the secretion of specific cargo proteins or cell wall components. Plant cytoskeleton influences the dynamics of exocytosis through various functions. First, secretory vesicles are delivered near the plasma membrane. Second, microtubules were shown to mark the place of exocytosis. Third, cytoskeleton is able to prevent membrane fusion by simple separation of compartments. Fourth, cytoskeleton potentially mediates the interaction between molecules of secretory apparatus. Secretion of certain cargo molecules appears to be dependent on different cytoskeleton types and the exocytosis seems to be specifically regulated in each tissue. This thesis aims to describe interplay of cytoskeleton and secretory pathway on the example of tip growth and to predict future direction of research on secretory pathway based on cellulose synthase secretory data.
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