Bear Shields of Plains Indians : Motifs and Meanings

The study is the outcome of two research projects on the North American Indian traditions: the role of the shields within the Plains Indians traditional culture and religion, and the bear ceremonialism of the Native North America, especially the significance of the bear among the Plains Indians. This article-based dissertation includes seven separately published scholar papers, forming Chapters 6 12. The introduction formulates the objectives and frame of reference of the study and the conclusions pulls together its results. The study reconsiders the role of the Plains Indian shields with bear motifs. Such shields are found in rock art, in the Plains Indian s paintings and drawings, and in various collections, the main source material being the shields in European and North American museums. The aim is not only to study shields with bear power motifs and the meanings of the bear, but also to discuss appropriate methods for studying these subjects. There are three major aims of the study: to consider methodical questions in studying Plains Indian shields, to examine the complexity of the Plains Indian shields with the bear power motifs, and to offer new interpretations for the basic meanings of the bear among the Plains Indians and the interrelationship between individualism and collectivism in the Plains Indians visionary art that show bear power motifs on the shields. The study constructs a view on the bear shields taking account of all sources of information available and analysing the shields both as physical artefacts and religious objects from different perspectives, studying them as a part of the ensemble of Plains culture and religious traditions. The bear motifs represented the superhuman power that medicine men and warriors could exploit through visions. For the Plains Indians, the bear was a wise animal from which medicine men could get power for healing but also a dangerous animal from which warriors could get power for warfare. The shields with bear motifs represented the bear powers of the owners of the shields. The bear shield was made to represent the vision, and the principal interpretation of the symbolism was based on the individual experience of spiritual world and its powers. The study argues that the bear shield as personal medicine object is based on wider tribal traditions, and the basic meaning is derived from the collective tradition. This means that the bear seen in vision represented particular affairs and it was represented on the shield surface using conventional ways of traditional artistry. In consequence of this, the bear shields reflect not only the individual experiences of bear power but whole field of tribal traditions that legitimated the experiences and offered acceptable interpretations and conventional modes for the bear symbols.

Ivanov-mistik : Okkul'tnye motivy v poèzii i proze Vjačeslava Ivanova (1907-1919)

The subject of this work is the mysticism of Russian poet, critic and philosopher Vjacheslav Ivanov (1866-1949). The approach adopted involves the textual and discourse analysis and findings of the history of ideas. The subject has been considered important because of Ivanov's visions of his dead wife, writer Lydia Zinovieva-Annibal, which were combined with audible messages ("automatic writings"). Several automatic writings and descriptions of the visions from Ivanov's archive collections in St.Petersburg and Moscow are presented in this work. Right after the beginning of his hallucinations in the autumn of 1907, Ivanov was totally captivated by the theosophical ideas of Anna Mintslova, the background figure for this work. Anna Mintslova, a disciple of Rudolf Steiner's Esoteric School, offered Ivanov the theosophical concept of initiation to interpret paranormal phenomena in his intimate life. The work is divided into three main chapters, an introduction and aconclusion. The first chapter is called The Mystical Person: Anthropology of Ivanov and describes the role of the inner "Higher Self" in Ivanov's views on the nature of human consciousness. The political implications of the concepts, "mystical anarchism" and "sobornost" (religious unity) are also examined. The acquaintance and contacts with Anna Mintslova during 1906-1907 gave a framework to Ivanov's search for an organic society and personal religious experience. The second part, Mystics of Initiation and Visionary Aesthetics describes the influence of the initiation concept on Ivanov's aesthetic views (mainly "realistic symbolism"). On the other hand, some connections between the imagery of his visions and symbols in his verses of that period are established. Since Mintslova represented the ideas of Rudolf Steiner in Russia, several symbols shared by Steiner and Ivanov ("rose", "rose and cross") have been another subject of investigation. The preference for strict verse form in the lyrics of Ivanov's visionary period is interpreted as an attempt to place his own poetic creation within two traditions, a mystical and literary one. The third part of this work, Mystics of Hope and Terror, examines Ivanov's conception of Russia in connection with Mintslova's ideas of occult danger from the East. Ivanov's view of the "Russian idea" and his nationalistic idea during World War I are considered as a representation of the fear of the danger. Ivanov's interpretation of the October revolution is influenced by the theosophical concept of the "keeper of the threshold" which occurs in the context of the discourse of occult danger.

Ruumiillinen esine, materiaalinen suku : Tutkimus Pyhä Anna itse kolmantena -aiheisista keskiajan puuveistoksista Suomessa

Embobied Object, Material Family. Late-Medieval Wood Sculptures Depicting Saint Anne with the Virgin and Child in Finland Saint Anne, the mother of the Virgin Mary, was one of the most popular saints in Western Europe in the late Middle Ages. She was often depicted with two other figures, the Virgin and the Christ Child (Anna Selbdritt). The dissertation examines the polychrome wood sculptures showing this motif, with a special focus on those remaining in Finland. It investigates the meanings these sculptures had to their observers in the fifteenth-century Finland. The study sheds light to important material heritage which is little known and offers new insights into the cult and imagery of the holy grandmother. Methodologically the study is based on iconology and post-formalist art history, and it appropriates concepts such as spatiality, sanctity, corporeality, and gender. Taking a comparative approach it knits together larger tendencies and local people and incidents. By conflating methodological domains it renews the ways how fragmentary wood sculptures, lacking documentary written sources, can be contextually interpreted and comprehended. The sculptures are analyzed from three angles. Firstly, the study explores the sculptures by focusing on their materiality and facture, which is to consider them as records of their own making. The analysis provides new information concerning the quantity, location, and current condition of the sculptures and it also elucidates problems regarding attribution, dating, display, and craftsmanship. The book presents the results of the empirical study of 45 Saint Anne groups; these works are individually described in the large Appendix. Secondly, the works are contextualized to the specific historical conditions in which they were observed. The study discusses closely the circumstances in the Turku Cathedral around the shrine of Saint Anne, the popular belief, and the piety of individual persons. The sculptures, deemed as the embodiments of the holy characters, interacted with the devotees. Thirdly, the works are examined within the wider theological and ideological currents of the era centered on the body and Incarnation. Saint Anne with the Virgin and Child motif demonstrated the Carnal Trinity, the motherly side of the Holy Trinity. The dissertation argues that Saint Anne was interpreted as the female counterpart or, in a mythical sense, wife of God. Furthermore, the Child s implicit, simultaneous presence as a suffering or dead man imbues the sculptures with a sense of the Passion, thus associating them with the pietà and the Mater dolorosa motifs. The naked Christ Child underlines him as the offering and, eventually, the Eucharistic wafer. The study suggests that the sculptures mediate continuity and the bloodline between the generations by the intertwined and repeated gestures, clothing and positions of the portrayed figures. Regardless of the ostensible homeliness of the sculptures, so readily reiterated by earlier scholars, these sculptures represented creation and birth through the carnal yet holy mothers.

Kansallista vai modernia : Taidegrafiikka osana 1930-luvun taidejärjestelmää

The theme of this doctoral thesis is the Finnish printmaking in the years 1930-1939. During this decade, there were approximately 100 artists making prints in Finland. Indeed, the period was an especially important one for printmaking. Associations for printmakers were founded in Helsinki and Turku, training in the field was launched, and the number of printmaking exhibitions increased considerably. Through their national organisations, Finnish printmakers participated in many exhibitions abroad, interaction with Nordic printmakers being especially intense. Thus, a firm basis for post-war developments was created. However, printmakers' activity- which had continued throughout the 1930s - declined notably after the Winter War broke out in the autumn of 1939. As a result, the period 1930-1939 forms a coherent and distinct unity in Finnish printmaking history. The study consists of two parts: the main text and an appendix in which the production of each printmaking artist active in the 1930s is examined separately. The study also includes a comprehensive list of the prints made in the course of the decade. One of the central themes is the printmakers' relationship to "Finnish nationalist" art and concepts of art in the 1930s. I analyse the various manifestations of this way of thinking in the visual arts of the period. Finnish fine art in the period between the world wars has usually been characterised as conservative, introverted and spiritually isolated from the modern European trends of the time. On the basis of this study, such a view is too simple. Many artists and printmakers adopted a modernistic notion of art that approached the newest in European modernism, including such trends as avant-garde classicism and general European new Objective Realism (Die neue Sachlichkeit). On the other hand, choosing Finnish nationalist motifs did not necessarily mean that the artist was opposed to modernism: modernist artists could still be interested in national themes. The relationship of 1930s printmaking to the world of nationalist ideas is examined in this doctoral thesis from several perspectives. Towards the end of the main text, I examine the issue from the point of view of selected artists. Another feature that emerged during the study and turned out to be surprisingly widespread was the close relationship of many artists to religious, theosophical and pantheistic views. I deal with this issue in greater detail through a few representative printmakers.

Arkeen pudonnut sibylla : Modernin naisen identiteetin rakentuminen Marja-Liisa Vartion romaanissa Kaikki naiset näkevät unia

A Sibyl fallen into everyday life. The enfolding of the identity of modern woman in Marja- Liisa Vartio s novel Kaikki naiset näkevät unia ( All Women Have Dreams ). --- Marja-Liisa Vartio played a remarkable part in renewing Finnish literature. My thesis examines her novel Kaikki naiset näkevät unia (1960), which describes the life of a middle-aged housewife, Mrs. Pyy ( Mrs. Hazel Hen ). She has moved from country to city and lives now in a suburb, in the Helsinki of the 1950 s. In Finnish literature, the novel is the first significant description of a modern city woman accomplished by modernistic means. My research examines the identity of a woman in the Finland of the 50 s, an epoch marked by the inevitable transition into modernity. My aim is to look into the ways in which the female identity enfolds in Kaikki naiset näkevät unia, how it takes its form, how it is described and commented. The primary method is contextual close reading; the novel is seen in the social, cultural and historical context of the time it was published. Essential elements in this study are literary motifs and images in the novel, and particularly transtextual relations as defined by Gérald Genette. The focus is on hypertextuality, intertextuality and paratextuality. Kaikki naiset näkevät unia emerges as a modern version of Madame Bovary by Gustave Flaubert. A woman s life spent in illusive dreaming is transferred from a 1900th century bourgeois town in France to a middle class Finnish suburb in the 1950 s. Vartio s novel is a variant of an ancient Finnish ballad I, a bird without a nest , making it into a modern narration of transition. The inner, mental journey from country to city is of great length, and the liminal life in a suburb does not make the passage any easier. Like the lyrical voices in the poetry of Edith Södergran, also Mrs. Pyy finds it hard to discover any values of sisterhood or those of ideal femininity in modern times. In earlier studies of Marja-Liisa Vartio s prose, stress has been laid on the discourse of her narrators and characters, as well as on its modern literary form. In this research, however, urgent allusions to paintings, old and new, are taken into account, since Mrs. Pyy mirrors herself against art, both classical and modern. Principal images in this context are Michelangelo s Sibyls in the ceiling of the Sistine Chapel, and a modern painting, which remains unidentified. Mrs. Pyy turns out to be a tragicomic character, who has magnanimous illusions about herself, but is compelled to accept the fact the she is only a mediocre person. She is nothing more than a first generation city dweller; she is not a modern, aloof outsider but a mere dilettante, who desperately tries to live out modern city life. Kaikki naiset näkevät unia is a striking picture of the 1950 s, a picture that is construed in the consciousness of Mrs. Pyy. We are shown everyday life growing more and more modern after the war and woman s role growing more and more subject to increasing pressure for change.

A Touch of Red : Archaeological and Ethnographic Approaches to Interpreting Finnish Rock Paintings

Approximately 125 prehistoric rock paintings have been found in the modern territory of Finland. The paintings were done with red ochre and are almost without exception located on steep lakeshore cliffs associated with ancient water routes. Most of the sites are found in the central and eastern parts of the country, especially on the shores of Lakes Päijänne and Saimaa. Using shore displacement chronology, the art has been dated to ca. 5000 – 1500 BC. It was thus created mainly during the Stone Age and can be associated with the so-called ‘Comb Ware’ cultures of the Subneolithic period. The range of motifs is rather limited, consisting mainly of schematic depictions of stick-figure humans, elks, boats, handprints and geometric signs. Few paintings include any evidence of narrative scenes, making their interpretation a rather difficult task. In Finnish archaeological literature, the paintings have traditionally been associated with ’sympathetic’ hunting magic, or the belief that the ritual shooting of the painted animals would increase hunting luck. Some writers have also suggested totemistic and shamanistic readings of the art. This dissertation is a critical review of the interpretations offered of Finnish rock art and an exploration of the potentials of archaeological and ethnographic research in increasing our knowledge of its meaning. Methods used include ’formal’ approaches such as archaeological excavation, landscape analysis and the application of neuropsychological research to the study of rock art, as well as ethnographically ’informed’ approaches that make use of Saami and Baltic Finnish ethnohistorical sources in interpretation. In conclusion, it is argued that although North European hunter-gatherer rock art is often thought to lie beyond the reach of ‘informed’ knowledge, the exceptional continuity of prehistoric settlement in Finland validates the informed approach in the interpretation of Finnish rock paintings. The art can be confidently associated with shamanism of the kind still practiced by the Saami of Northern Fennoscandia in the historical period. Evidence of similar shamanistic practices, concepts and cosmology are also found in traditional Finnish-Karelian epic poetry. Previous readings of the art based on ‘hunting magic’ and totemism are rejected. Most of the paintings appear to depict experiences of falling into a trance, of shamanic metamorphosis and trance journeys, and of ‘spirit helper’ beings comparable to those employed by the Saami shaman (noaidi). As demonstrated by the results of an excavation at the rock painting of Valkeisaari, the painted cliffs themselves find a close parallel in the Saami cult of the 'sieidi', or sacred cliffs and boulders worshipped as expressing a supernatural power. Like the Saami, the prehistoric inhabitants of the Finnish Lake Region seem to have believed that certain cliffs were ’alive’ and inhabited by the spirit helpers of the shaman. The rock paintings can thus be associated with shamanic vision quests, and the making of ‘art’ with an effort to socialize the other members of the community, especially the ritual specialists, with trance visions. However, the paintings were not merely to be looked at. The red ochre handprints pressed on images of elks, as well as the fact that many paintings appear ’smeared’, indicate that they were also to be touched – perhaps in order to tap into the supernatural potency inherent in the cliff and in the paintings of spirit animals.

Playing "Nordic": The Women of Niskavuori, Agri/Culture, and Imagining Finland on the Third Reich Stage

The theatrical censorship of the Third Reich considered the playwright's race and politics alongside the content of the drama. Given the political stigma of its "leftist" author, it is rather surprising that Hella Wuolijoki's Niskavuoren naiset opened in 1938 at the Staatliches Schauspielhaus in Hamburg. The play ran for fourteen performances before being closed by the Reichsdramaturgie, apparently at the instigation of Finnish critics. Yet this was not the end of the play's or its author's fortunes in the Third Reich, as the possibility of staging the play was raised several times over the next four years, coming to a close in 1942. Playing "Nordic" examines the ideological and theatrical background of this extended "cultural performance," as a means to reopening and reconstructing the work of the 1938 Die Frauen auf Niskavuori. Written by a Finnish, northern, "Nordic" author, and preoccupied with the dynamics of rural culture in an increasingly urbanized world, Niskavuoren naiset was understood in the Third Reich to illustrate and reinforce the racial, agri/cultural themes of Blut und Boden ("veri ja maa"). Playing "Nordic" examines this thematic relationship in three phases. The first phase uses archival materials to investigate the Reichsdramaturgie's understanding of the play and its author, and its ongoing discussion of Wuolijoki from 1937 to 1942. Play evaluator Sigmund Graff's description of Niskavuoren naiset as hamsunartig, or "Hamsun-esque," inspires the second phase of the dissertation, which first elaborates the meanings of Blut und Boden through a reading of contemporary "racial" theory and anthropology, and then assesses the representation of Finland within this discourse, one of the dominant cultural paradigms of the Third Reich. Imaging Finland for German audiences, the play stood among analogous, continued efforts to represent Finland and the rural life in the Third Reich, colored by Blut und Boden: art and agricultural exhibitions, essays and propaganda literature, mass demonstrations of the peasantry. This wider framework for the performance of "Finland" materializes the abstract or theoretical program of Blut und Boden in its everyday performed meanings; as such it provides the essential background for reading the Hamburg production of Die Frauen auf Niskavuori, which sustains the third and final phase. The German translation and the Hamburg photographic record are compared with the Helsinki premiere to assess the impact of Blut und Boden on the representation of Wuolijoki's play in the Third Reich. The journalistic critical response illuminates the effect that the dramatic complex of rural and racial values - generically identified as Bauerndrama in the Third Reich - had on the reception of the play; at the same time, both visual and critical documents also suggest possible moments of theatrical dissent in the Hamburg production. Playing "Nordic" undertakes a documentary and cultural reading of the changing theatrical meanings of Wuolijoki's Niskavuoren naiset as it crossed the frontier from Finland to the stage of the Third Reich. It also provides a model for the ways theatrical signification operates within a network of cultural and ideological meanings, suggesting the ideological work of theatrical production depends on, reinforces, and contests that tissue of values. Although Finnish criticism of Niskavuoren naiset has assumed the play's Blut und Boden resonance contributed to Wuolijoki's success in the Third Reich, this study shows a considerably more complex situation. This revealing production dramatizes the changing uses of plays in a politicized national and transnational context. As part of the framing of "Nordic" identity on the wider stage of the Third Reich, Die Frauen auf Niskavuori exemplifies the conjunction of concurrent - sometimes independent, sometimes interlocking - "racial" and national ideologies.

New Perspectives on the Roman Coinage on the Eastern Limes in the Late Republican and Roman Imperial Period

The Ph.D. thesis discusses the monetary development in Roman Syria and Judaea in the Late Republican and the Early Imperial Period, from a numismatic, archaeological and historical point of view. In effect, the work focuses on the 1st century B.C. to the 1st century A.D., that is, the assumed time of introduction of Roman denarii to the region. The work benefits from the silver coin hoards of Khirbet Qumran recently published by the author. Though discovered as early as 1955 at Qumran, where the famous Dead Sea Scrolls had been found prior to that in 1947, most hoards remained unpublished until 2007. A second important source utilized is the so-called Tax Law from Palmyra in Syria. Its significance lies in the fact that Palmyra used to be one of the most important cities on the Silk Road, along which luxury goods were transported into the Roman Empire and Rome itself. During the research conducted, studies of the provincial coinage of Judaea (A.D. 6-66) shed new light on the authority of the Roman governors in economic and monetary matters in eastern Mediterranean regions. Furthermore, a new suggestion as to the length of the mandate period of Pontius Pilate is made. The extent of Emperor Augustus monetary reforms as well as the military history of Judaea are discussed in the light of new analytical studies, which show that the production of Roman base metal coins appears to have been a highly controlled process, contrary to popular opinion. Statistical calculations related to the coin alloy revealed striking similarities with Roman and other local metalwork found in Israel; a fact previously unknown. Results indicate that both Roman and local metalwork consisted of outstandingly systematized practises and may have exploited the same metal sources. Information: Kenneth Lönnqvist (*25.7.1962) has studied at the University of Helsinki since 1981. Furthermore, Lönnqvist has lived in the Mediterranean countries and the Near East, and made research there at various scientific institutions and universities for ca. 7 years. Contact and sales of thesis: kenneth.lonnqvist@helsinki.fi

Viisi vuotta tiedonrakentamista yhteisö- ja teknologiaelementtien muovautumisen näkökulmasta

The object of this study was to examine the phenomena of a long-term Knowledge Building process. The subject was OECD/ENSI/FI-project's Knowledge Building in Knowledge Forum®3.4 environment from 8.9.2000 to 8.9.2005. Research was based on socio-cognitive and socio-cultural learning approaches and the theoretical background consisted of models of collaborative learning and knowledge processing. These theoretical applications were first structured using metaphors of language and then assembled into five main theoretical motifs. The main motifs were 1) context, 2) inter-subjective, shared area, 3) community's practices and participation, 4) developing expertise and 5) the sequential construction of processes. These themes were assembled in interpreting the results using the Mutual Shaping of Technological and Social Elements by Boczkowski (1999) as a conceptual tool. The social elements of the mutual shaping process were defined as 1) community structure, 2) discourse and 3) the meanings of activity. The technological elements were defined as 1) shared artefacts, 2) features of technology-use and 3) other technological conventions perceived in activity. The five main theoretical motifs were used as the basis for creating the research problems, which were divided into three themes: 1) shared artefacts, themes of Knowledge Building and participant formation, 2) patterns of participation and interaction and 3) the meanings of activity. As methods I used content analysis of the messages, the quantitative profiling of changes in the database, social network analysis, discourse analysis of selected message threads and theme interviews of eleven participants. Based on my study it's possible to say, that a long-term setting of this kind provides a different perspective on Knowledge Building from most of the previous research. The most valuable conclusions from the data are: 1) The centralisation of interaction in this type of setting is a feature that supports the improvement in the quality of action. 2) The participation in a long-term Knowledge Building process seems to support the concious effort on professional development and the expert-identity. 3) The quality of plasticity of the technology-in-use has implication for how the communal features of activity will develop. The agency is seen to initiate processes that in turn open up new possibilities for the quality of action on both the communal and individual levels.

Molecular Details of Serum Resistance of Yersinia enterocolitica

In complement activation, Factor H (FH) and C4b-binding protein (C4bp) are the key regulators that prevent the complement cascade from attacking host tissues. Some bacteria may bind and deposit these regulators on their own surfaces and thus provide themselves with an efficient means to avoid complement activation. In consequence, bacteria resist complement-mediated lysis and opsonin-dependent phagocytosis. This study has demonstrated that Y. enterocolitica, similar to many other pathogens, recruits both FH and C4bp to its surface to ensure protection against the complement-mediated killing. YadA and Ail, the most crucial serum resistance factors of Y.enterocolitica, mediate the binding of FH and C4bp. FH - YadA interaction involves multiple higher structural motifs on the YadA stalk and the short consensus repeats (SCRs) of the entire polypeptide chain of FH. The Ail binding site on FH has been located to SCRs 6 and 7. The binding site for FH on Ail, however, remains undetermined. Both YadA- and Ail-bound regulators display full cofactor activity for FI-mediated cleavage of C3b/C4b. FH/C4bp-binding characteristics do, however, differ between YadA and Ail. In addition, Ail captures the regulators only in the absence of blocking lipopolysaccharide O-antigen and outer core, whereas YadA binds FH/C4bp independent of the presence of other surface factors Independent of mode of binding, however, YadA and Ail provide Y. enterocolitica a means to avoid complement-mediated lysis, enhancing chances for the bacteria to survive in the host during various phases of infection.

ORP1L, a new Rab7 effector and regulator of late endosome functions

Oxysterol binding protein (OSBP) homologues have been found in eukaryotic organisms ranging from yeast to humans. These evolutionary conserved proteins have in common the presence of an OSBP-related domain (ORD) which contains the fully conserved EQVSHHPP sequence motif. The ORD forms a barrel structure that binds sterols in its interior. Other domains and sequence elements found in OSBP-homologues include pleckstrin homology domains, ankyrin repeats and two phenylalanines in an acidic tract (FFAT) motifs, which target the proteins to distinct subcellular compartments. OSBP homologues have been implicated in a wide range of intracellular processes, including vesicle trafficking, lipid metabolism and cell signaling, but little is known about the functional mechanisms of these proteins. The human family of OSBP homologues consists of twelve OSBP-related proteins (ORP). This thesis work is focused on one of the family members, ORP1, of which two variants were found to be expressed tissue-specifically in humans. The shorter variant, ORP1S contains an ORD only. The N-terminally extended variant, ORP1L, comprises a pleckstrin homology domain and three ankyrin repeats in addition to the ORD. The two ORP1 variants differ in intracellular localization. ORP1S is cytosolic, while the ankyrin repeat region of ORP1L targets the protein to late endosomes/lysosomes. This part of ORP1L also has profound effects on late endosomal morphology, inducing perinuclear clustering of late endosomes. A central aim of this study was to identify molecular interactions of ORP1L on late endosomes. The morphological changes of late endosomes induced by overexpressed ORP1L implies involvement of small Rab GTPases, regulators of organelle motility, tethering, docking and/or fusion, in generation of the phenotype. A direct interaction was demonstrated between ORP1L and active Rab7. ORP1L prolongs the active state of Rab7 by stabilizing its GTP-bound form. The clustering of late endosomes/lysosomes was also shown to be linked to the minus end-directed microtubule-based dynein-dynactin motor complex through the ankyrin repeat region of ORP1L. ORP1L, Rab7 and the Rab7-interacting lysosomal protein (RILP) were found to be part of the same effector complex recruiting the dynein-dynactin complex to late endosomes, thereby promoting minus end-directed movement. The proteins were found to be physically close to each other on late endosomes and RILP was found to stabilize the ORP1L-Rab7 interaction. It is possible that ORP1L and RILP bind to each other through their C-terminal and N-terminal regions, respectively, when they are bridged by Rab7. With the results of this study we have been able to place a member of the uncharacterized OSBP-family, ORP1L, in the endocytic pathway, where it regulates motility and possibly fusion of late endosomes through interaction with the small GTPase Rab7.

Bending the Rules of Cell Protrusions : Molecular Mechanisms and Biological Roles of Inverse-BAR Proteins in Cell Morphogenesis

Plasma membrane adopts myriad of different shapes to carry out essential cellular processes such as nutrient uptake, immunological defence mechanisms and cell migration. Therefore, the details how different plasma membrane structures are made and remodelled are of the upmost importance. Bending of plasma membrane into different shapes requires substantial amount of force, which can be provided by the actin cytoskeleton, however, the molecules that regulate the interplay between the actin cytoskeleton and plasma membrane have remained elusive. Recent findings have placed new types of effectors at sites of plasma membrane remodelling, including BAR proteins, which can directly bind and deform plasma membrane into different shapes. In addition to their membrane-bending abilities, BAR proteins also harbor protein domains that intimately link them to the actin cytoskeleton. The ancient BAR domain fold has evolved into at least three structurally and functionally different sub-groups: the BAR, F-BAR and I-BAR domains. This thesis work describes the discovery and functional characterization of the Inverse-BAR domains (I-BARs). Using synthetic model membranes, we have shown that I-BAR domains bind and deform membranes into tubular structures through a binding-surface composed of positively charged amino acids. Importantly, the membrane-binding surface of I-BAR domains displays an inverse geometry to that of the BAR and F-BAR domains, and these structural differences explain why I-BAR domains induce cell protrusions whereas BAR and most F-BAR domains induce cell invaginations. In addition, our results indicate that the binding of I-BAR domains to membranes can alter the spatial organization of phosphoinositides within membranes. Intriguingly, we also found that some I-BAR domains can insert helical motifs into the membrane bilayer, which has important consequences for their membrane binding/bending functions. In mammals there are five I-BAR domain containing proteins. Cell biological studies on ABBA revealed that it is highly expressed in radial glial cells during the development of the central nervous system and plays an important role in the extension process of radial glia-like C6R cells by regulating lamellipodial dynamics through its I-BAR domain. To reveal the role of these proteins in the context of animals, we analyzed MIM knockout mice and found that MIM is required for proper renal functions in adult mice. MIM deficient mice displayed a severe urine concentration defect due to defective intercellular junctions of the kidney epithelia. Consistently, MIM localized to adherens junctions in cultured kidney epithelial cells, where it promoted actin assembly through its I-BAR andWH2 domains. In summary, this thesis describes the mechanism how I-BAR proteins deform membranes and provides information about the biological role of these proteins, which to our knowledge are the first proteins that have been shown to directly deform plasma membrane to make cell protrusions.

Folding and Selective Exit of Reporter Proteins from the Yeast Endoplasmic Reticulum

The present study analyses the traffic of Hsp150 fusion proteins through the endoplasmic reticulum (ER) of yeast cells, from their post-translational translocation and folding to their exit from the ER via a selective COPI-independent pathway. The reporter proteins used in the present work are: Hsp150p, an O-glycosylated natural secretory protein of Saccharomyces cerevisiae, as well as fusion proteins consisting of a fragment of Hsp150 that facilitates in the yeast ER proper folding of heterologous proteins fused to it. It is thought that newly synthesized polypeptides are kept in an unfolded form by cytosolic chaperones to facilitate the post-translational translocation across the ER membrane. However, beta-lactamase, fused to the Hsp150 fragment, folds in the cytosol into bioactive conformation. Irreversible binding of benzylpenicillin locked beta-lactamase into a globular conformation, and prevented the translocation of the fusion protein. This indicates that under normal conditions the beta-lactamase portion unfolds for translocation. Cytosolic machinery must be responsible for the unfolding. The unfolding is a prerequisite for translocation through the Sec61 channel into the lumen of the ER, where the polypeptide is again folded into a bioactive and secretion-competent conformation. Lhs1p is a member of the Hsp70 family, which functions in the conformational repair of misfolded proteins in the yeast ER. It contains Hsp70 motifs, thus it has been thought to be an ATPase, like other Hsp70 members. In order to understand its activity, authentic Lhs1p and its recombinant forms expressed in E. coli, were purified. However, no ATPase activity of Lhs1p could be detected. Nor could physical interaction between Lhs1p and activators of the ER Hsp70 chaperone Kar2p, such as the J-domain proteins Sec63p, Scj1p, and Jem1p and the nucleotide exchange factor Sil1p, be demonstrated. The domain structure of Lhs1p was modelled, and found to consist of an ATPase-like domain, a domain resembling the peptide-binding domain (PBD) of Hsp70 proteins, and a C-terminal extension. Crosslinking experiments showed that Lhs1p and Kar2p interact. The interacting domains were the C-terminal extension of Lhs1p and the ATPase domain of Kar2p, and this interaction was independent of ATPase activity of Kar2p. A model is presented where the C-terminal part of Lhs1p forms a Bag-like 3 helices bundle that might serve in the nucleotide exchange function for Kar2p in translocation and folding of secretory proteins in the ER. Exit of secretory proteins in COPII-coated vesicles is believed to be dependent of retrograde transport from the Golgi to the ER in COPI-coated vesicles. It is thought that receptors escaping to the Golgi must be recycled back to the ER exit sites to recruit cargo proteins. We found that Hsp150 leaves the ER even in the absence of functional COPI-traffic from the Golgi to the ER. Thus, an alternative, COPI-independent ER exit pathway must exists, and Hsp150 is recruited to this route. The region containing the signature guiding Hsp150 to this alternative pathway was mapped.