Functional role of the Mso1p-Sec1p complex in membrane fusion regulation

Sec1/Munc18 (SM) protein family members are evolutionary conserved proteins. They perform an essential, albeit poorly understood function in SNARE complex formation in membrane fusion. In addition to the SNARE complex components, only a few SM protein binding proteins are known. Typically, their binding modes to SM proteins and their contribution to the membrane fusion regulation is poorly characterised. We identified Mso1p as a novel Sec1p interacting partner. It was shown that Mso1p and Sec1p interact at sites of polarised secretion and that this localisation is dependent on the Rab GTPase Sec4p and its GEF Sec2p. Using targeted mutagenesis and N- and C-terminal deletants, it was discovered that the interaction between an N-terminal peptide of Mso1p and the putative Syntaxin N-peptide binding area in Sec1p domain 1 is important for membrane fusion regulation. The yeast Syntaxin homologues Sso1p and Sso2p lack the N-terminal peptide. Our results show that in addition to binding to the putative N-peptide binding area in Sec1p, Mso1p can interact with Sso1p and Sso2p. This result suggests that Mso1p can mimic the N-peptide binding to facilitate membrane fusion. In addition to Mso1p, a novel role in membrane fusion regulation was revealed for the Sec1p C-terminal tail, which is missing in its mammalian homologues. Deletion of the Sec1p-tail results in temperature sensitive growth and reduced sporulation. Using in vivo and in vitro experiments, it was shown that the Sec1p-tail mediates SNARE complex binding and assembly. These results propose a regulatory role for the Sec1p-tail in SNARE complex formation. Furthermore, two novel interaction partners for Mso1p, the Rab GTPase Sec4p and plasma membrane phospholipids, were identified. The Sec4p link was identified using Bimolecular Fluorescence Complementation assays with Mso1p and the non-SNARE binding Sec1p(1-657). The assay revealed that Mso1p can target Sec1p(1-657) to sites of secretion. This effect is mediated via the Mso1p C-terminus, which previously has been genetically linked to Sec4p. These results and in vitro binding experiments suggest that Mso1p acts in cooperation with the GTP-bound form of Sec4p on vesicle-like structures prior to membrane fusion. Mso1p shares homology with the PIP2 binding domain of the mammalian Munc18 binding Mint proteins. It was shown both in vivo and in vitro that Mso1p is a phospholipid inserting protein and that this insertion is mediated by the conserved Mso1p amino terminus. In vivo, the Mso1p phospholipid binding is needed for sporulation and Mso1p-Sec1p localisation at the sites of secretion at the plasma membrane. The results reveal a novel layer of membrane fusion regulation in exocytosis and propose a coordinating role for Mso1p in connection with membrane lipids, Sec1p, Sec4p and SNARE complexes in this process.

ISO 13567 - The Proposed International Standard for Structuring Layers in Computer Aided Building Design

Layering is a widely used method for structuring data in CAD-models. During the last few years national standardisation organisations, professional associations, user groups for particular CAD-systems, individual companies etc. have issued numerous standards and guidelines for the naming and structuring of layers in building design. In order to increase the integration of CAD data in the industry as a whole ISO recently decided to define an international standard for layer usage. The resulting standard proposal, ISO 13567, is a rather complex framework standard which strives to be more of a union than the least common denominator of the capabilities of existing guidelines. A number of principles have been followed in the design of the proposal. The first one is the separation of the conceptual organisation of information (semantics) from the way this information is coded (syntax). The second one is orthogonality - the fact that many ways of classifying information are independent of each other and can be applied in combinations. The third overriding principle is the reuse of existing national or international standards whenever appropriate. The fourth principle allows users to apply well-defined subsets of the overall superset of possible layernames. This article describes the semantic organisation of the standard proposal as well as its default syntax. Important information categories deal with the party responsible for the information, the type of building element shown, whether a layer contains the direct graphical description of a building part or additional information needed in an output drawing etc. Non-mandatory information categories facilitate the structuring of information in rebuilding projects, use of layers for spatial grouping in large multi-storey projects, and storing multiple representations intended for different drawing scales in the same model. Pilot testing of ISO 13567 is currently being carried out in a number of countries which have been involved in the definition of the standard. In the article two implementations, which have been carried out independently in Sweden and Finland, are described. The article concludes with a discussion of the benefits and possible drawbacks of the standard. Incremental development within the industry, (where ”best practice” can become ”common practice” via a standard such as ISO 13567), is contrasted with the more idealistic scenario of building product models. The relationship between CAD-layering, document management product modelling and building element classification is also discussed.

Reacting to Complex Threats – the Securitization in the Speeches of Anders Fogh Rasmussen based on NATO´s New Strategic Concept

The purpose of this master´s thesis is to analyze how NATO Secretary General Anders Fogh Rasmussen is trying to justify the existence of the military alliance through the use of security arguments. I am puzzled by the question: why does NATO still exist – what is NATO’s raison d'être. The New Strategic Concept (2010) forms the base for his argumentation. This thesis focuses on the security argumentation of NATO which is examined by analyzing the speeches the Secretary General. The theoretical framework of this study is based on constructivist approach to international security examining the linguistic process of securitization. Issues become securitized after Anders Fogh Rasmussen names them as threats. This thesis focuses on the securitization process relating to NATO and analyses what issues Rasmussen raises to the security agenda. Research data consists of the speeches by Anders Fogh Rasmussen. They are analyzed through J.L. Austin’s speech act taxonomy and Chaïm Perelman’s argumentation theories. The thesis will concentrate on the formulation and articulation of these threats which are considered and coined as “new threats” in contemporary international relations. I am conducting this research through the use of securitization theory. This study illustrates that the threats are constructed by NATO’s member-states in unison, but the resolutions are sounded through Rasmussen’s official speeches and transcripts. . Based on the analysis it can be concluded that Rasmussen is giving reasons for the existence of NATO. This takes place by making use of speech acts and different rhetorical techniques. The results of the analysis indicate that NATO remains an essential organization for the West and the rest of the world according to the Secretary General.

“And now to the rest of the trash…” Representations of begging Roma and an ideal society in two Nordic newspapers

Since begging East European Roma became a common view in the streets of larger Nordic cities, vivid discussions about their presence and activities have been carried out in the mass media. This thesis examines the public debates in Finland and Norway through a discursive analysis and comparison of press content from the two countries. The aim of the study is firstly to identify the prominent discourses which construct certain images of the beggars, as well as the elements and internal logics that these discourses are constructed around. But in addition to scrutinizing representations of the Roma, also an opposite perspective is applied. In accordance with the theoretical concept of ‘othering’, debates about ‘them’ are assumed to simultaneously reveal something significant about ‘us’. The second research question is thus what kind of images of the ideal Finnish and Norwegian societies are reflected in the data, and which societal values are salient in these images. The analysis comprises 79 texts printed in the main Finnish and Norwegian quality newspapers; Helsingin Sanomat and Aftenposten. The data consists of news articles, editorials, columns and letters to the editor from a three-month period in the summer of 2010. The analysis was carried out within the theoretical and methodological framework of critical discourse analysis as outlined by Norman Fairclough. A customized nine-step coding scheme was developed in order to reach the most central dimensions of the texts. Seven main discourses were identified; the Deprivation-solidarity, Human rights, Order, Crime, Space and majority reactions, Authority control, and Authority critique discourse. These were grouped into two competing normative stances on what an ideal society looks like; the exclusionary and the inclusionary stance. While the exclusionary stance places the begging Roma within a frame of crime, illegitimate use of public space and threat to the social order, the other advocates an attitude of solidarity and humanitarian values. The analysis points to a dominance of the former, although it is challenged by the latter. The Roma are “individualized” by quoting and/or presenting them by name in a fair part of the Finnish news articles. In Norway, the opposite is true; there the beggars are dominantly presented as anonymous and passive. Overall, the begging Roma are subjected to a double bind as they are faced with simultaneous expectations of activity and passivity. Theories relating to moral panics and ‘the good enemy’ provide for a deepened understanding of the intensity of the debates. Keywords: East European Roma, begging, media, newspapers, Helsingin Sanomat, Aftenposten, critical discourse analysis, Norman Fairclough, othering, ideal society, moral panics, good enemy, double bind, Finland, Norway