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Actin and the ARP 2/3 complex in the nucleus
Němcová, Barbora ; Bellinvia, Erica (advisor) ; Hála, Michal (referee)
The eukaryotic actin cytoskeleton is required for numerous cellular processes, including cell shape, development and movement, gene expression and signal transduction, and response to biotic and abiotic stress. Actin constitutes a wide family of proteins that are major components of the cytoskeleton. Actin is one of the most abundant proteins in living organisms. Actin has essential functions both in the cytoplasm and in the nucleus, where it has been linked to key nuclear processes. Recent studies have shown that actin is actively transported from the cytoplasm to the nucleus, where it regulates transcriptional aktivity, regulates RNA polymerases, is involved in chromatin remodeling and repair damaged DNA. The presence of typical actin filaments in the nucleus has not been demonstrated directly.but nuclear actin occurs in many forms such as actin rods, short actin polymers, actin monomers, or actin complexes with profilin or cofilin. Most eukaryotic cells also contain at least eleven actin-related proteins (ARPs). Although many ARPs are cytoskeletal, recent biochemical and genetic work has demonstrated that some ARPs function largely or entirely in the nucleus. Nuclear ARPs are recognized as novel key regulators of genome function, and affect not only the remodeling of chromatin but also the...
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Functions of actin and myosin 1c in the cell nucleus and in the cytoplasm
Kalendová, Alžběta ; Hozák, Pavel (advisor) ; Binarová, Pavla (referee) ; Forstová, Jitka (referee)
Human MYO1C gene encodes three myosin 1c (Myo1c) isoforms which differ only at their N-ends. Interestingly, all three isoforms localize to the nucleus and also to the cytoplasm, where they are anchored to the plasma membrane by the interaction with phosphatidyl inositol-4,5-bisphosphate (PIP2). However, studies reporting functional involvement of these isoforms are inconsistent. While the shortest isoform C (Myo1c-isoC) has been implicated exclusively in the cytoplasmic processes, the longer isoform B (termed the nuclear myosin 1, NM1) has been employed in the nuclear and processes, such as DNA transcription and rRNA maturation. Similarly, the longest isoform A (Myo1c-isoA) exerts its functions in the nucleus solely. To complete the information on the cellular functions of Myo1c isoforms, we searched for the cytoplasmic functions of NM1 and nuclear functions of Myo1c-isoC. In mouse, only two isoforms (NM1 and Myo1c-isoC) are expressed. We prepared the knock-out mouse (KO) which lacks specifically NM1 while retaining Myo1c-isoC unchanged. Surprisingly, this manifested in no phenotype observed. Since we demonstrated that even Myo1c-isoC acts in the transcription in the similar manner as NM1, it suggests that Myo1c- isoC functionally overlap with NM1 in the nuclear functions. Besides its localization...
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The function of direct forms of presentation in the generic structure of newspaper reports
Urbanová, Zuzana ; Dušková, Libuše (advisor) ; Malá, Markéta (referee) ; Hornová, Libuše (referee)
The thesis deals with the employment of direct forms of presentation in the genre of hard news. The texts for the analysis were excerpted from the main British broadsheet newspapers. The classification of forms of presentation is based on Semino and Short (2004) and focuses on their deictic and syntactic properties, interpreted in terms of the pragmatic concepts of perspective, faithfulness claims and the role of the reported and reporting speaker. Attention is paid to various direct forms, including direct speech, free direct speech and combined forms, i.e. non-direct forms appearing with a partial direct quote. The thesis draws heavily on the work by White (1998), whose approach to genre is informed by the ideas proposed by the Sydney School and Systemic Functional Linguistics. Hard news is characterised by the orbital generic structure, consisting of the nucleus and a number of specifying satellites (White 1998). The occurrence of direct and combined forms of presentation is explained by their deictic, syntactic and pragmatic properties, the generic role and characteristic features of the nucleus and individual satellites, and the overall function of hard news. The function of hard news and forms of presentation is also discussed in terms of the more general concepts of heteroglossia, dialogue...
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Formy a funkce přímé řeči v žánrové struktuře novinových zpráv
Urbanová, Zuzana ; Dušková, Libuše (advisor) ; Hornová, Libuše (referee) ; Malá, Markéta (referee)
Zuzana Urbanová The Function of Direct Forms of Presentation in the Generic Structure of Newspaper Reports Abstract The thesis deals with the employment of direct forms of presentation in the genre of hard news. The texts for the analysis were excerpted from the main British broadsheet newspapers. The classification of forms of presentation is based on Semino and Short (2004) and focuses on their deictic and syntactic properties, interpreted in terms of the pragmatic concepts of perspective, faithfulness claims and the role of the reported and reporting speaker. Attention is paid to various direct forms, including direct speech, free direct speech and combined forms, i.e. non-direct forms appearing with a partial direct quote. The thesis draws heavily on the work by White (1998), whose approach to genre is informed by the ideas proposed by the Sydney School and Systemic Functional Linguistics. Hard news is characterised by the orbital generic structure, consisting of the nucleus and a number of specifying satellites (White 1998). The occurrence of direct and combined forms of presentation is explained by their deictic, syntactic and pragmatic properties, the generic role and characteristic features of the nucleus and individual satellites, and the overall function of hard news. The function of hard news and...
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Nuclear dynamics and interactions of myosin 1c
Dzijak, Rastislav ; Hozák, Pavel (advisor) ; Hašek, Jiří (referee) ; Kaňka, Jiří (referee)
1. ABSTRACT Myosins are proteins that convert chemical energy stored in ATP into mechanical force that is applied on an actin filament. Nuclear myosin 1 (NM1) was the first myosin detected in the cell nucleus. Together with nuclear actin they were shown to play important roles in DNA transcription and chromatin remodeling. However, the molecular details of the NM1 functions are largely unknown. To expand our knowledge about this molecular motor we studied tissue expression, mechanism of nuclear localization and molecular interactions of this myosin motor. In the first part we examined the expression pattern of NM1 in various mouse tissues. We demonstrated that NM1 is present in cell nuclei of all mouse tissues examined except for cells in terminal stages of spermatogenesis. Quantitative PCR and western blots demonstrated that the expression of NM1 in tissues varies, with the highest levels in the lungs. NM1 is a nuclear isoform of earlier identified myosin 1c (Myo1c), which was described initially as a cytosolic, and plasma membrane associated protein. The only known difference between these two proteins was the presence of additional 16 amino acids at the N-terminus of NM1. Next we focused on the influence of NM1 domains, including the N-terminus, on the subcellular localization of this protein. We found...
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Giardia intestinalis karyotypes
Hudosová, Lenka ; Nohýnková, Eva (advisor) ; Král, Jiří (referee)
Giardia intestinalis is a parasitic protist that causes one of the most common diarrheal disease of parasite origin. The cell of Giardia contains two nuclei with unknown number of chromosomes until recently. Karyotype was determined five years ago using conventional cytogenetic method by Tůmová and collaborators. In my work, I assessed karyotype of four isolates, six lines and three clonal lines by the same method. It was confirmed, that two nuclei within one cell could differ in chromosome number, the differences found were 1, 2 or 6 chromosomes. Aneuploid number of chromosomes was found too. In case that both nuclei within single cell contained the same number of chromosomes, there were 10 chromosomes indentified in each nucleus. It was also revealed, that karyotype is not specific feature for different genetic groups (in this work assemblages A and E). Karyotype can be different even among lines and clonal populations derived from the same isolate. Changes in karyotype in the course of in vitro cultivation were detected within three populations. Results are discussed in relation to known facts.
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The role of tubulin transport between the nucleus and the cytoplasm
Bokvaj, Pavel ; Schwarzerová, Kateřina (advisor) ; Zažímalová, Eva (referee)
Tubulin is the basic building block of microtubules which ensure intracellular transport, morphological changes, the connection of proteins of metabolic pathways, the segregation of chromosomes during mitosis and many other essential processes in plant cells. The localization of tubulin regarding nucleus is strictly controlled and during the interphase, tubulin is usually not present in the nucleus. As we previously showed, one of the exceptions of the presence of tubulin in the nucleus is the cold stress condition. Also, we discovered several plant-specific nuclear export sequences (NES) in tubulin molecules: one in α-tubulin and two in β-tubulin. In this work I found that double mutation of both functional β-tubulin NESes cause slight accumulation of the protein in the nuclei. However, the phenotype of the transgenic plants expressing β-tubulin with mutations is probably not affected by the presence of the mutated protein. The effect of the expression of the mutated β-tubulins was observed in Nicotiana tabacum BY-2 cells as well. The expression of β-tubulin carrying a single NES3 mutation or double mutation of both NESes bring changes of the division activity of the cells, while the NES2 mutation does not have any effect. Furthermore, it was discovered that the 0řC cold tr eatment does not cause...
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Photon strength functions in 162Dy from two-step gamma cascade measurement
Chudoba, Petr ; Krtička, Milan (advisor) ; Knapp, František (referee)
Photon strength functions quantities, which describe deexcitation of nucleus by emiting gamma rays within statistical model of the nucleus. Information about these quantities can be gained from study of two-step gamma cascades. The thesis is focused on processing of experimental data from measurement of these cascades in 162Dy nucleus. Data were acquired using experimental set-up istalled on the research reactor in Řež near Prague. In the experiment the 161Dy targed was bombarded by thermal neutrons. Following gamma deexcitation of 162Dy nucleus was measured by coincident spectrometer composed of two semi-conductor detectors. The aim of the thesis is to perform energy and efficiency calibration and to process experimental data into the form of two-step gamma cascades spectra. In adition, a comparsion of the two-step cascades spectra with the outcome of simulations of a few basic models of photon strength functions is made.
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