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Dynamics of ALBA proteins in Arabidopsis thaliana evaluated by fluorescence microscopy
Popelářová, Anna ; Honys, David (advisor) ; Vosolsobě, Stanislav (referee)
ALBA proteins were discovered in Archaea more than 30 years ago. They were gradually identified to be well conserved in Eucaryotes as well. A functional dimeric form of these proteins with DNA and RNA-binding capability was claimed in both mentioned domains of organisms. However, their roles diversified during evolution and vary in between organisms. In Archaea, ALBAs are involved in the genome organization and RNA-protein interactions. In Eukaryotes, there are presented two different subfamilies of ALBA proteins - Rpp20 and Rpp25 subfamily. A sole protein from each subfamily was identified in some organisms though they were multiplied in plants, respectively. These proteins can interact with each other and participate in ontogenetic development and stress responses. According to several studies, ALBA proteins were found to be involved in DNA stability maintenance or pre-rRNA splicing in the nucleus of Arabidopsis thaliana. However, they have been shown to play a role in the cellular metabolism and stress responses in cytoplasm. Six ALBA proteins were identified in the genome of A. thaliana, three from each subfamily. In this study, all heterodimeric protein- protein interactions were investigated by the bimolecular fluorescence complementation (BiFC) assay which revealed positive results in...
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Functional characterization of Alba-family genes in Arabidopsis thaliana
Kočová, Helena ; Honys, David (advisor) ; Fischer, Lukáš (referee)
(anglicky) Alba-family proteins were identified in Archaea and Eucarya and are classified among the oldest and the most conserved nucleic acid-binding proteins. The binding preferences and roles differ among certain evolution clades. In Crenarchaea they represent chromatin-binding proteins, while their role in RNA metabolism is suggested in Euryarchaea and Eukaryotes. ALBA proteins are well characterized in human, where they play a role in the RNAse P/MRP complex and in unicellular parasites, such as Plasmodium and Trypanosoma, where an involvement in the life cycle regulation is confirmed. In plants, their role is not yet well understood. The aim of this thesis is to increase a knowledge about the Alba-family proteins in the model plant Arabidopsis thaliana. Based on a minimal changes to development and reproduction in single mutants and high sequence similarity, a functional redundancy of the proteins was assumed. For better understanding of the ALBA proteins function, three smaller members of the family were edited by the same metod. The obtained triple mutant showed delay in flowering. ALBA dimer formation was confirmed in many organisms. BiFC method was used to determine Arabidopsis ALBA homodimerization. The data analysis showed potential homodimerization in most of them.
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