Interleukin-1alpha (IL-1alpha) is a well-known proinflammatory mediator acting as a secreted molecule. However, in addition to its ability to activate its membrane-bound receptor, there is growing evidence on its noncanonical nuclear function, which classifies IL-1alpha as a "dual function cytokine". This nuclear action depends on the evolutionary conserved N-terminal domain of IL-1alpha. After proteolytic processing, the N-terminal domain of IL-1alpha translocates into nucleus. Histone acetyltransferase (HAT) complexes were previously identified as nuclear targets of IL-1alpha precursor. However, the specific protein which is responsible for the interaction between IL-1alpha and HAT complexes has not been identified yet. To dissect this interaction, the N-terminal domain of IL-1alpha was produced in yeast. Suitability of this experimental setup for testing the interaction between IL-1alpha and eukaryotic HAT complexes was evaluated in this study. IL-1alpha has been analyzed in this study using bioinformatics approaches as well. Putative amphipatic acidic helixes of IL-1alpha have been characterized. One of the potential binding partners of these domains is protein Ada2. Protein Ada2, mature IL-1alpha and IL-1alpha precursor in fusion with epitopes suitable for affinity purification were produced in...

Mitochondrien haben eine entscheidende Rolle im Zellmetabolismus, da sie den Hauptort der ATP-Produktion darstellen. Störungen des mitochondrialen Metabolismus sind mit einem weiten Spektrum von Erkrankungen assoziiert. Das Gehirn und die Muskulatur sind aufgrund ihres hohen Energiebedarfs dabei oft betroffen (Epilepsie, Ataxie, Myopathie). Diese Arbeit beschreibt die Klonierung von nukleären Genen des Komplexes I der mitochondrialen Atmungskette. Besonderes Augenmerk wird dabei auf die 51 kDa Untereinheit (NDUFV1) gerichtet, da sie mit ihrer Bindungsstelle für NADH2 die Eintrittspforte in den Komplex I darstellt. In dieser Untereinheit werden die ersten Mutationen beschrieben, die bei Kindern zu schwerer Entwicklungsretardierung, Leukenzephalopathie und Muskelhypotonie führen. Im weiteren werden Patienten mit isoliertem Komplex III Mangel molekulargenetisch untersucht und klassifiziert. Bei einem Patienten war ein isolierter Komplex III-Mangel und eine Mutation im mitochondrialen Cytochrom b-Gen mit einer septo-optischen Dysplasie vergesellschaftet. Am Ende beschreibt die Arbeit die Probleme der pränatalen Diagnostik mitochondrialer Erkrankungen und die Besonderheiten der genetischen Beratung betroffener Familien. / Mitochondria have a crucial role in the energy metabolism of the cell, since they constitute the main place for ATP-production. Defects in the mitochondrial metabolism are associated with a wide spectrum of diseases. Due to their high energy demand brain and muscles are regularly affected (epilepsy, ataxia, myopathy). This work describes the cloning of nuclear encoded genes of complex I of the mitochondrial respiratory chain. The main interest is directed towards the 51 kDa subunit (NDUFV1) since, due to its NADH2-binding domain, it constitutes the entry port into complex I. Therein the first mutations are described, which lead to severe developmental delay, leukencephalopathy and muscular hypotonia in infants. Additionally patients with isolated complex III-deficiency are examined molecularly and are classified according to their clinical symptoms. In one patient isolated complex III deficiency and a mutation in the mitochondrial cytochrome b-gene are associated with septo-optic dysplasia. At the end problems with prenatal diagnosis of mitochondrial diseases and the peculiarities of genetic counselling of affected families are discussed.

The dynamics and function of the actin cytoskeleton depends on polymerization and branching of actin filaments, an event that is stimulated by Arp2/3. Arp2/3-dependent branching is closely linked to the pentameric WASH complex which consists of WASH, strumpellin, SWIP, CCDC53 and FAM21. WASH complex is associated mainly with endosomes. It was traditionally localized to retromer-coated domains of early endosomes which enable sorting and recycling of endocytosed material. However, latest scientific data extend the role of WASH complex to other endosomal or even non-endosomal sites. Of all the subunits of the WASH complex, FAM21 is the most prominent hub for protein-protein interactions, thanks to its long unstructured C-terminal domain. In my diploma thesis FAM21 was localized to early and late endosomes and lysosomes of U2OS human cell line. Dictyostelium discoideum was then used as a model organism to investigate FAM21 protein interactions as well as the proteins associated specifically with the C terminal domain of FAM21. Results of the study shed new light on the complex network of FAM21 interactions and question the long-standing theories on the function of WASH complex in cells. Powered by TCPDF (www.tcpdf.org)

This Master's thesis deals with diversity within the Micrasterias papillifera/ M. radiosa (Desmidiales) species complex. Both species are inhabitants of endangered biotops, like peat-bogs and, like other desmids, are used for biomonitoring. Many described varieties for both of them can be found in taxonomic literature, but their morphology is overlapping and revision of this species complex seemed to be necessary. All strains were isolated from peat-bogs and lakes across Europe and my dataset was properly completed by strains from algal collections. Results of phylogenetic analysis of sequences for ITS and psaA showed, that the real intraspecific diversity of Micrasterias papillifera Brébisson ex Ralfs was overestimated. Although I could find a little intraspecific diversity within the ‚papillifera' lineage in the phylogenetic tree based on ITS sequences, it did not correspond with a morphology of isolates. M. papillifera possess unexpected morphological plasticity and also some isolates designated as Micrasterias radiosa Ralfs were present in this lineage. On the contrary, there were only a few isolates in the lineage ‚radiosa'. This species seems to be relatively rare and probably has limited distribution. Both species are morphologically well delimited from each other on the basis of several characters...

During the transformation of the bacterial endosymbiont into current mitochondria the protein import apparatus had to be created de novo. The reduced mitochondria (mitosomes) of the parasitic protist Giardia intestinalis represent unique cellular model for the examination of these fundamental transport processes. As the main objective of this project I will try to characterize the motor complex, which propels the protein transport, and also the translocation channel in the inner mitosomal membrane. To this aim I will exploit the presence of two membrane components Pam16 and Pam18, which were discovered in our laboratory, and which constitute the functional core of the motor complex. Based on the information from the analogous systems of yeast and mammalian mitochondria, these two components should physically interact with so far unknown translocation channel. In all other eukaryotes this channel is formed by a conserved protein Tim23. The absence of this protein in the genome of G. intestinalis suggests presence of completely novel, or maybe the original-bacterial protein. Having in hand this simplified mitochodrial model the project has potential to bring not only new data in parasite biology but also generate new information on the function and evolution of mitochondrial protein import.

Current studies illustrate that the development of the controller role has been where its focusis more about acting in the managements interest and to be a "business partner" which createsmore value for the company. Since most studies are completed in the private sector, we wanted to take the opportunity to examine the public sector to see whether the development has been carried out there. A development of the controller role means that the controller will have wider duties which could lead to an overload and non-controllable role. We have studied both new and old articles, collected and compared controller's duties towards our empiricism. We chose to limit ourselves to four municipalities in Skaraborg where we conducted a total of eight interviews to draw the fairest possible analysis. The purpose of the study was to see how controllers and even municipalities have been affected by this development, as recent studies present. After having compiled our empirical data collection, we could conclude that the majority feel that the tasks have become wider and where the skills are also becoming increasingly important. In the current situation, the basis of our empirical data says that controllers have not been particularly affected by the development. Respondents feel that the controller role isunder control and not overloaded, but at the same time that the development must be slowed down. Otherwise it will not be long before such a situation arises. For this, we have also linked how important Controller's skills is to be able to handle this now so broad professional role. / Nuvarande studier belyser att en utveckling av controllerrollen har skett där dennes fokus alltmer handlar om att agera i ledningens intresse och vara en ”business partner” för att på så vis skapa mer värde åt företaget. Eftersom de flesta av dessa studier är gjorda inom den privata sektorn ville vi passa på att undersöka den offentliga sektorn för att se om samma utveckling har genomförts där. En utveckling av yrkesrollen innebär att controllern får bredare arbetsuppgifter vilket skulle kunna leda till att rollen blir överbelastad och icke-kontrollerbar. Vi har med hjälp av äldre artiklar kombinerat med nyare artiklar ställt och jämfört controllerns arbetsuppgifter mot vår empiri. Vi valde att avgränsa oss till fyra kommuner inom Skaraborg där vi genomförde totalt åtta kvalitativa intervjuer för att kunna dra en så rättvis analys som möjligt. Syftet med studien var att kunna se hur controllers och även kommunerna har påverkats av denna utveckling som nyare studier tar upp. Efter att ha sammanställt vår empiriinsamling kunde vi dra slutsatsen att majoriteten upplever att arbetsuppgifterna har blivit bredare och därav har även kompetensen blivit allt viktigare. I dagsläget kan vi inte utifrån vår empiri säga att controllers har påverkats särskilt mycket av utvecklingen. Respondenterna upplever att yrkesrollen är under kontroll och inte överbelastad, men samtidigt att utvecklingen måste bromsas. Annars dröjer det inte länge förrän en sådan situation uppstår. Till detta har vi även kopplat hur pass viktig controllerns kompetens är för att kunna hantera denna nu så breda yrkesroll.

This diploma thesis deals with the energy policy of the Russian Federation in the period from 2000 to 2008 and its prospects until 2030. First of all, I analyse particular targets of the energy policy of Russia and put them in a broader framework. Consequently, I focus my attention on three economic targets that need to be met if Russia is to become an "energetic superpower". Firstly, I search for the answer to the question whether Russia has enough oil and gas for foreigner markets and whether it can provide for their export in order to be able to act successfully as an "energetic superpower". Secondly, whether Russia will succeed when exporting oil and natural gas to world markets and whether it will manage to diversify transport routes for its exports. Thirdly, whether the effort to "make the economy move" supported by raw-material exports has been successful and whether this situation can continue. The thesis further follows each answer to these questions in two dimensions of time. A hindsight shall first clarify how successful the energy policy of Russia was when meeting these targets until 2008, or what problems Russia had to face while trying to meet them. In consequence, this provides a basis for a foresight.

Splicing is a process, during which non-coding sequences (introns) are cleaved out of pre-mRNA, and exons are ligated. This whole process is catalysed by a multi-megadalton splicing complex, composed of five small nuclear ribonucleoprotein particles (shortly snRNPs), which each contains its own small nuclear RNA molecule and specific set of proteins. During the biogenesis of snRNPs, U4 and U6 snRNPs are assembled to form the di-snRNP, which further associates with U5 snRNP and gives rise to tri-snRNP. With the help of mass spectrometry, we have found previously uncharacterized protein interacting with U5 snRNP, called TSSC4. By immunoprecipitation, I confirmed TSSC4 as a U5 snRNP specific protein and identified the region of TSSC4 responsible for interaction with U5 snRNP. I also showed that TSSC4 interacts with PRPF19, a component of complex driving the catalytic activation of the spliceosome and that this interaction is U5 snRNP-independent. Knockdown of TSSC4 in HeLa cells results in accumulation of di-snRNAs and U5 snRNP in Cajal bodies, nuclear compartments involved in snRNP biogenesis. Similar phenotype was previously observed upon inhibition of tri-snRNP assembly. To analyse the importance of TSSC4 for tri-snRNP assembly, I separated individual snRNPs by glycerol gradient ultracentrifugation...

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)...

No description available.