Münster (Westfalen), Univ., Diss., 2005 / Dateien im PDF-Format

Universiẗat, Diss., 2002--Bremen.

Universiẗat, Diss., 2002--Bremen.

Techn. University, Diss., 2005--Berlin.

No description available.

WASH complex regulates actin dynamics on endosomes by activating the Arp2/3 complex, which subsequently induces generation of branched actin patches. WASH complex is required for proper recycling of many important transmembrane proteins. Although the general physiological function of WASH complex is known, the role of its single subunits have not yet been adequately specified. This work focuses on one of these subunits - protein SWIP. This protein maintains vesicular localization of some WASH complex subunits in the slime mold Dictyostelium discoideum and a point mutation in its sequence causes a severe neurodegenerative disease - autosomal recessive intellectual disorder (ARID). Our results show that SWIP truncation results in its inability to incorporate into WASH complex. However, the C-terminal part of SWIP is able to localize onto intracellular vesicles, which are not WASH complex positive. We have also studied the impact of ARID-causing SWIP mutation, and we show, that it does neither change the protein's ability to bind the complex nor the overall localization of WASH complex.

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1 Abstract Even though mitochondria possess their own genome and ribosomes, majority of mitochondrial proteins is encoded in the nucleus and translated by cytosolic ribosomes. Hence it was necessary to establish transport complexes allowing the import of proteins from the cytosol. These complexes are best described in yeast. However, we are encountering organisms lacking many of the subunits of these complexes with increasing frequency. Therefore, we are presenting the overview of the distribution of the subunits within eukaryotic organisms. We specifically take a closer look at parasitic protist Giardia intestinalis that is well known for its extreme reductions of the import complexes. There have been only few subunits identified so far. Porin Tom40, that is responsible for translocating all the incoming proteins across the outer mitochondrial membrane, has been identified despite the high divergence, while homolog of Sam50 hasn't been successfully identified yet. Sam50 is however believed to be necessary for insertion of Tom40 into the membrane. Vast part of this thesis is dedicated to this phenomenon that is highly uncommon and maybe unique among the eukaryotic organisms.

The subject of the diploma thesis was the ideological elaboration of the architectural study of the building of the civilo temple. The proposal follows the elaborated urban concept of reclamation of the sub-castle - the territory of the historical Vydrica area and its surroundings. The building is a conceptual translation of the term profane sacrum into an architectural formalization. The main ideology consists in transforming the term profan sacrum into an object that incorporates three spirits - the spirit of the place, the spirit of the mind and the spirit of the heart. Thus, an artistic complex will be created on the historical basis of the center of gravity of Bratislava history. The spirit of the place is represented by the historical formalization of the base of the building which refers to the defunct Fish Square, the spirit of the mind represents a complex of 3 objects of self-development: object of performing arts, gallery and temple- library, and the last object which represents the spirit of the heart is transferred to the community center building. The overall ideology is to create a place that will be accessible every day and will serve to build our inner spirit. To develop internally and become a better person. A place to perceive, a place to develop oneself, a place to help each other, a place to share. A place where I will learn to play the violin and in the next phase I will organize a concert in the auditorium for the community until the phase where I will teach others. A place where I learn to cook and then sell my products on the market. A place to help each other. Better place for individuals, better place for community, better place for Bratislava.

Der Synaptonemalkomplex (SC) ist eine hochkonservierte Proteinstruktur. Er weist eine dreiteili-ge, leiterähnliche Organisation auf und ist für die stabile Paarung der homologen Chromosomen während der Prophase der ersten meiotischen Teilung verantwortlich, die auch als Synpase be-zeichnet wird. Fehler während der Synpase führen zu Aneuploidie oder Apoptose der sich entwi-ckelnden Keimzellen. Seit 1956 ist der SC Gegenstand intensiver Forschung. Seine Existenz wurde in zahlreichen Orga-nismen von der Hefe bis zum Menschen beschrieben. Seine Struktur aus zwei parallel verlaufen-den Lateralelementen (LE), die durch eine Vielzahl von sogenannten Transversalfilamenten (TF) verbunden werden und dem Zentralen Element (CE) in der Mitte des SC ist dabei offensichtlich über die Millionen von Jahren der Evolution erhalten geblieben. Einzelne Proteinkomponenten des SC wurden jedoch nur in wenigen Modelorganismen charakterisiert, darunter Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster, Ceanorhabditis elegans und Mus mus-culus. Unerwarteter Weise gelang es bei dieser Charakterisierung nicht, eine evolutionäre Ver-wandtschaft, d.h. eine Homologie zwischen den Proteinsequenzen der verschiedenen SCs nach-zuweisen. Diese Tatsache sprach gegen die grundsätzliche Annahme, dass der SC in der Evolution nur einmal entstanden sei. Diese Arbeit hat sich nun der Aufgabe gewidmet, die Diskrepanz zwischen der hochkonservierten Struktur des SC und seiner augenscheinlich nicht-homologen Proteinzusammensetzung zu lösen. Dabei beschränkt sie sich auf die Analyse des Tierreichs. Es ist die erste Studie zur Evolution des SC in Metazoa und demonstriert die Monophylie der Säuger SC Proteinkomponenten im Tierreich. Die Arbeit zeigt, dass mindestens vier von sieben SC Proteinen der Maus spätestens im letzten gemeinsamen Vorfahren der Gewebetiere (Eumetazoa) enstanden sind und auch damals Teil ei-nes ursprünglichen SC waren, wie er heute in dem Nesseltier Hydra zu finden ist. Dieser SC weist die typische Struktur auf und besitzt bereits alle notwendigen Komponenten, um die drei Domä-nen – LE, TF und CE – zu assemblieren. Darüber hinaus ergaben die einzelnen Phylogenien der verschiedenen SC Proteine der Maus, dass der SC eine sehr dynamische Evolutionsgeschichte durchlaufen hat. Zusätzliche Proteine wurden während der Entstehung der Bilateria und der Wir-beltiere in den SC integriert, während andere ursprüngliche Komponenten möglicherweise Gen-Duplikationen erfuhren bzw. besonders in der Linie der Häutungstiere verloren gingen oder sich stark veränderten. Es wird die These aufgestellt, dass die auf den ersten Blick nicht-homologen SC Proteine der Fruchtfliege und des Fadenwurms tatsächlich doch von den ursprünglichen Prote-inenkomponenten abstammen, sich aber aufgrund der rasanten Evolution der Arthropoden und der Nematoden bis zu deren Unkenntlichkeit diversifizierten. Zusätzlich stellt die Arbeit Hydra als alternatives wirbelloses Modellsystem für die Meiose- und SC-Forschung zu den üblichen Modellen D. melanogaster und C. elegans vor. Die kürzlich gewon-nenen Erkenntnisse über den Hydra SC sowie der Einsatz der Standard-Methoden in diesem Orga-nismus werden in dem abschließenden Kapitel zusammengefasst und diskutiert. / The synaptonemal complex (SC) is a highly conserved structure in sexually reproducing organism. It has a tripartite, ladder-like organization and mediates the stable pairing, called synapsis, of the homologous chromosomes during prophase of meiosis I. Failure in homolog synapsis result in aneuploidy and/or apoptosis of the developing germ cells. Since 1956, the SC is subject of intense research and its presence was described in various species from yeast to human. Its structure was maintained during millions of years of evolution consist-ing of two parallel lateral elements (LEs), joined by numerous transverse filaments (TFs) which run perpendicular to the LEs and an electron dense central element (CE) in the middle of the SC. Individual protein components, however, were characterized only in few available model organ-isms, as for example Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster, Ceanorhabditis elegans and Mus musculus. Rather unexpectedly, these characterizations failed to detect an evolutionary homology between the protein components of the different SCs. This fact challenged the general idea of a single origin of the SC in the evolution of meiosis and sexual reproduction. This thesis now addressed itself to the task to unravel the discrepancy between the high conser-vation of the SC structure and its diverse and apparently non-homologous protein composition, focusing on the animal kingdom. It is the first study dealing with the evolution of the SC in Meta-zoa and demonstrates the monophyly of the mammalian SC components in metazoan species. The thesis demonstrates that at least four out of seven murine SC proteins emerged in Eumeta-zoa at the latest and have been likewise part of an ancient SC as it can be found in the present-day cnidarian species Hydra. This SC displays the common organization and already possesses the minimal protein kit corresponding to the three different structural domains: LEs, TFs and the CE. Additionally, the individual phylogenies of the murine SC proteins revealed the dynamic evolu-tionary history of the ancient SC. Further components were added during the diversification of Bilateria and vertebrates while ancestral proteins likely duplicated in the vertebrate lineage and diversified or got lost in the branch leading to ecdysozoan species. It is hypothesized that the apparently non-homologous SC proteins in D. melanogaster and C. elegans actually do derive from the ancient SC proteins but diversified beyond recognition during the fast evolution of Ar-thropoda and Nematoda. The study proposes Hydra as an alternative invertebrate model system for meiosis and SC re-search to the standard organisms D. melanogaster and C. elegans. Recent results about the cni-darian SC as well as the possible application of standard methods is discussed and summarized in the concluding section.