|
|
Functional studies of selected members of the Arabidopsis formin family
Oulehlová, Denisa ; Cvrčková, Fatima (advisor) ; Binarová, Pavla (referee) ; Černý, Jan (referee)
Formins are multidomain proteins containing a conserved formin-homology 2 (FH2) domain, which catalyzes de novo nucleation of actin filaments. In yeast and animal cells, both mechanisms and regulation of formin function have been extensively studied, yet much less is known about action of plant formins, which considerably differ from yeast and animal ones in the domain composition. In higher plants, formins are classified into two groups, Class I and Class II, and so far, experimental data are available only for the first group members. Here I present results of experimental study of several members of the large formin family in Arabidopsis, including the characterization of a Class II formin AtFH16. Arabidopsis genome contains 21 formin-encoding genes, and though they greatly differ in their expression levels and pattern, all of them are transcriptionally active. We selected 17 homozygous T-DNA insertional mutants in 14 formin genes. Under standard cultivation conditions, no obvious phenotypic discrepancies between wild type and mutant plants were found. To impair two dominant pollen formins, an atfh3atfh5 double-mutant was prepared and even in this case, both microspore development and pollen tube growth remained unaffected. Consistently, polarized growth of tobacco pollen tubes was not altered...
|
|
|
Pharmacologically active compounds in hops
Pýchová, Kateřina ; Cvrčková, Fatima (advisor) ; Mašková, Petra (referee)
The hop plant (Humulus lupulus) is used not only in brewing but also in phytopharmacology and phytotherapy. Among the isolated or chemically produced substances used for healing are xanthohumol, 8-prenylnaringenine and α and β-bitter acids. Nowadays, a lot of attention is paid to their effect on humans - especially to their anticancer activity. But there is a lack of information about the importance of these substances for the hop plant itself. Most of the plant secondary metabolites originally serve for protection of the plant against pathogens and herbivores. The selected metabolites are not an exception. The purpose of this work was to summarize the published data about the biological importance of selected hop secondary metabolites and the effects they have on other organisms than plants, with an emphasis on humans.
|
|
|
Analysis of WASH complex member strumpellin
Pácalt, Ondřej ; Libusová, Lenka (advisor) ; Cvrčková, Fatima (referee)
Actin polymerization facilitated by the Arp2/3 complex plays a critical role in a wide range of cellular processes such as motility, endocytosis and cargo recycling. Activation and appropriate localization of the Arp2/3 complex is mediated by an interaction with the nucleation-promoting factor (NPF). WASH complex is the major endosomal NPF which plays a crucial role in the cargo recycling back to the trans-Golgi network (TGN) or plasma membrane. It is composed of five subunits: WASH1, SWIP, FAM21, CCDC53 and strumpellin. While WASH1 and FAM21 have been extensively studied, much less is known about strumpellin, a protein causally implicated in the onset of hereditary spastic paraplegia (HSP). This work focuses on strumpellin function in the cells, showing that only full-length protein incorporates into the WASH complex. In a strumpellin knock out cell line, we demonstrated that loss of strumpellin resulted in destabilization of the other WASH complex subunits. Still, an incomplete WASH complex without strumpellin was assembled. Cells also displayed enlarged endosomal subdomains and WASH complex nucleation activity on endosomes was largely diminished as assessed by loss of the actin patches. Finally, the absence of strumpellin was also accompanied by the accumulation of glucose transporter 1 (GLUT1)...
|
|
|
Plant tubulin code
Ničová, Klára ; Schwarzerová, Kateřina (advisor) ; Cvrčková, Fatima (referee)
Microtubules, which are made of polymers of α- and β-tubulin, are an integral part of the cytoskeleton. Both types of tubulin share a considerable sequential homology across eukaryotic organisms. Tubulins are encoded by relatively large gene families. The expression of these genes produces different tubulin isotypes, some of which may exhibit different properties. Tubulin isotypes can be further posttranslationally modified. The best known posttranslational modifications of tubulin include acetylation, phosphorylation, tyrosination, polyglutamylation and polyglycylation. The tubulin code arises from the combination of expression of different tubulin isotypes and their posttranslational modifications. As a consequence, microtubules in cells can be composed of a "mixture" of different tubulins with distinct functions and properties. Even though the existence of the tubulin code has been proven in every model organism, plants included, the precise understanding of the meaning of microtubules being composed of different tubulins is still subjected to research. Much of the research on the tubulin code has been carried out on animal models. In contrast, relatively little is known about the existence of the tubulin code in plants. This theses summarizes current knowledge on the localization and regulation...
|
|
| |
|
| |
|
|
Analysis of WASH complex member strumpellin
Pácalt, Ondřej ; Libusová, Lenka (advisor) ; Cvrčková, Fatima (referee)
Actin polymerization facilitated by the Arp2/3 complex plays a critical role in a wide range of cellular processes such as motility, endocytosis and cargo recycling. Activation and appropriate localization of the Arp2/3 complex is mediated by an interaction with the nucleation-promoting factor (NPF). WASH complex is the major endosomal NPF which plays a crucial role in the cargo recycling back to the trans-Golgi network (TGN) or plasma membrane. It is composed of five subunits: WASH1, SWIP, FAM21, CCDC53 and strumpellin. While WASH1 and FAM21 have been extensively studied, much less is known about strumpellin, a protein causally implicated in the onset of hereditary spastic paraplegia (HSP). This work focuses on strumpellin function in the cells, showing that only full-length protein incorporates into the WASH complex. In a strumpellin knock out cell line, we demonstrated that loss of strumpellin resulted in destabilization of the other WASH complex subunits. Still, an incomplete WASH complex without strumpellin was assembled. Cells also displayed enlarged endosomal subdomains and WASH complex nucleation activity on endosomes was largely diminished as assessed by loss of the actin patches. Finally, the absence of strumpellin was also accompanied by the accumulation of glucose transporter 1 (GLUT1)...
|
|
|
Comparative phenotypic study of selected Arabidopsis formin mutants
D'Agostino, Viktoria ; Cvrčková, Fatima (advisor) ; Tylová, Edita (referee)
Actin filaments and microtubules are involved in cell development and morphogenesis. Plant Class II formins regulate both cytoskeletal polymers. However their function has not yet been fully described. This study examines effects of LOF mutations in Arabidopsis thaliana FH13 (AT5G58160) and FH14 (AT1G31810) genes on early root system development using a pharmacological approach. Since measuring root length of numerous mutant lines in multiple conditions is laborious and time consuming, this thesis also involves optimization of this process with the aim to establish a reliable method of fast visualisation and measurement of Arabidopsis seedlings in a time series in the laboratory. Furthermore, statistical analysis for a large amount of data gathered in multiple conditions had to be optimized. While no significant phenotype in terms of root length was found in fh13, fh14 and double fh13 fh14 LOF mutants under standard conditions, treatment with cytoskeletal drugs revealed possible changes in lateral root branching in an fh14 mutant. Nevertheless, specific function of FH13 and FH14 remains a question.
|
|
| |
|
|
Transcriptional response of plants to genotoxic stress and water deficit
Libus, Jiří ; Štorchová, Helena (advisor) ; Cvrčková, Fatima (referee) ; Kovařík, Aleš (referee)
7 Conclusions 1) cDNA array was prepared and used to assay transcript abundances of 376 selected Ara- bidopsis transcripts following various treatments with MMS. a) LoTrEC, clustering algorithm based on local trends of expression profiles, was de- signed and applied to the data. It succeeded to discover functionally rellevant clusters of expression profiles. b) Transcriptional responses to various MMS treatment regimes were investigated. While high MMS concentration seemed only to induce nonspecific stress reaction, the low and combined MMS resulted in a set of more specific expression changes. c) Expression levels of five transcripts were estimated by qRT-PCR. Trends of most of the profiles were confirmed. 2) Expression of 3 genes related to drought stress and/or response to cytokinin were mea- sured by qRT-PCR in wild type and ZOG1 transgenic plants. Transcript levels of all the genes were altered by water deficit. a) Although there are no significant macroscopic differences between wild type and ZOG1 transgenic plants, the mRNA abundances appeared to be influenced by the genotype. b) Leaf position (age) significantly influenced the expression of cig1 and ZOG1 driven by SAG12 promoter. c) RT primed with oligo-dT appeared more eficient than random hexanucleotide-primed reaction for 3 out of 5 mRNAs...
|
guest ::
