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Proteinová rodina HSP70 a její role v biotických interakcích
Čičmanec, Petr
Heat shock proteins 70 (HSP70s) are ubiquitous and widespread proteins across prokaryotic and eukaryotic organisms, and accumulated evidence indicates that these proteins are involved in much more than the response to heat shock. This thesis characterizes the role of HSP70s in response to different stimuli and provides insight into the HSP70's role in regulating the interaction between biotic and abiotic factors. The theoretical part summarizes the current knowledge of HSP70, including information about its structure, activity, posttranslational modifications, and the so-called chaperone code. It describes the functions of HSP70s and pays particular attention to the HSP70s' role in immunity and biotic interactions. The experimental part is divided into several sections. First, a bioinformatic analysis provides a cross-kingdom comparison of HSP70 and its role in biotic stress. This part is supplemented with the analysis of HSP70 distribution and abundance at the proteome level. Next, the effect of different stimuli on HSP70 protein abundance in different model organisms is presented, including E. coli, S. cerevisiae, A. thaliana, Pisum sativum, and Linum usitatissimum. Finally, the effects of the hsp70 mutation are analyzed, providing insight into the complexity of the HSP70 machinery and confirming its role in the biotic interaction between a model plant and fungi.
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Studying dimer formation and effectors of Arabidopsis thaliana nascent polypeptide-associated complex
Klodová, Božena ; Fíla, Jan (advisor) ; Robert Boisivon, Helene (referee)
The development of plant flowers represents a complex process controlled by numerous mechanisms. The creation of double homozygous mutant of both β subunits (sometimes also referred to as basic transcription factor 3) of nascent polypeptide associated complex in Arabidopsis thaliana (further referred to as nacβ1 nacβ2) caused quite a strong defective phenotype including abnormal number of flower organs, shorter siliques with a reduced seed set, and inferior pollen germination rate together with a lower ovule targeting efficiency. Previously, NAC complex was described to be formed as a heterodimer composed of an α- and β-subunit, which binds ribosome and acts as a chaperone in Saccharomyces cerevisiae. In plants, NACβ is connected to stress tolerance and to plant development as a transcription regulator. However, little is known of NAC heterodimer function in plants. In this thesis, yeast two hybrid system (Y2H) and bimolecular fluorescence complementation (BiFC) assays were used to verify the NAC heterodimer formation in A. thaliana and to establish any potential interaction preferences between both NACβ paralogues and five NACα paralogues. To deepen the understanding about molecular mechanisms behind the nacβ1 nacβ2 phenotype, flower bud transcriptome of the nacβ1 nacβ2 double homozygous mutants...
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