The Krüppel-associated box (KRAB) zinc finger protein (ZFP) genes comprise one of the largest gene families in the mammalian genome, encoding transcription factors with an N-terminal KRAB domain and C-terminal zinc fingers. The KRAB domain interacts with a co-repressor protein, KAP-1, which can recruit various factors causing transcriptional repression of genes to which KRAB ZFPs bind. Little is currently known about the gene targets of the ~400 human and mouse KRAB ZFPs. Many KRAB ZFPs interact with factors other than KAP-1. To identify proteins that may interact with one particular KRAB ZFP, Zfp647, I previously carried out a yeast two-hybrid screen using the full-length Zfp647 sequence and a mouse embryonic cDNA library. I have now tested the interactions from this screen for their specificity for Zfp647. I show that Zfp647 can interact with itself and at least 20 other KRAB ZFPs through their zinc finger domains, and have confirmed the Zfp647 self-interaction by in vitro co-immunoprecipitation. In my yeast two-hybrid screen, Zfp647 bound to KAP-1 as well as another related protein, ARD1/Trim23. Zfp647 also interacts with proteins that function in ubiquitylation. I have found evidence to suggest that Zfp647 may also interact with proteins encoding jumonji domains both by yeast two-hybrid assay and by co-immunoprecipitation from NIH/3T3 cell extracts. We have previously found that Zfp647 localises to non-heterochromatic nuclear foci in differentiated ES cells, which also contain KAP-1 and HP1, and which lie adjacent to PML nuclear bodies in a high proportion of cells. I have found that these foci are also visible in pMEFs, but not NIH/3T3 tissue culture cells. Immunofluorescence studies with antibodies against proteins from the yeast twohybrid screen have not shown any significant co-localisation with Zfp647. KAP-1 is sumoylated ex vivo, as are two human KRAB ZFPs. Because Zfp647 lies adjacent to PML nuclear bodies and can associate with proteins involved in posttranslational modification, I tested whether Zfp647 is also modified. I characterised a sheep _-Zfp647 antibody previously created in the lab and have shown that it detects Zfp647 by western blot, but not by immunofluorescence. I show that treatment of NIH/3T3 cells with NEM, which prevents the removal of protein modifications, leads to the appearance of higher molecular weight forms of Zfp647. Modification of Zfp647 is not dependent on KAP-1, which is known to function as a SUMO E3 ligase. Attempts to classify the modification as either ubiquitin, SUMO or NEDD8 have suggested that Zfp647 may be mono-ubquitylated. The larger modified forms of Zfp647 are present in both NIH/3T3 and ES cells. Interestingly, I found that the modification profile of the protein changes over the course of ES cell differentiation, during which time Zfp647 relocalises to punctate nuclear foci; thus Zfp647 modification may be involved in this process.

In this project a theory of adaptive differential insurgency growth by the mechanism of repression driven contagion is put forth to explain variation in the membership and spatial expansion of insurgencies from 1981 to 1999. As an alternative to the dominant structural approaches in the civil war literature, Part 1 of the study proposes an interactive model of insurgency growth based on Most and Starr's opportunity and willingness framework. The findings suggest that state capacity, via its impact on state repressive behavior, plays an important gatekeeping function in selecting which minor insurgencies can grow into civil war, but contributes little to insurgency growth directly. In Part 2 of the study, I directly examine variation in insurgency membership and geographical expansion as a function of repression driven contagion. I find that repression increases the overall magnitude of insurgency activity within states, while at the same time reducing the density of insurgency activity in any one place. Despite an abundance of low intensity armed struggles against a highly diverse group of regimes around the world, I find an extremely strong and robust regularity: where repression is low - insurgencies don't grow.

Resistance to antiestrogen therapy remains a significant problem in breast cancer. Low expression of inhibitor of growth 4 (ING4) in primary tumors has been correlated with increased rates of recurrence in estrogen receptor-positive (ER+) breast cancer patients, suggesting a role for ING4 in ER signaling. This study provides evidence that ING4 inhibits ER activity. ING4 overexpression increased the sensitivity of T47D and MCF7 ER+ breast cancer cells to hormone deprivation. ING4 attenuated maximal estrogen-dependent cell growth without affecting the dose-response of estrogen. These results indicated that ING4 functions as a noncompetitive inhibitor of estrogen signaling and may inhibit estrogen-independent ER activity. Supportive of this, treatment with fulvestrant but not tamoxifen rendered T47D cells sensitive to hormone deprivation as did ING4 overexpression. ING4 did not affect nuclear ER alpha protein expression, but repressed selective ER-target gene transcription. Taken together, these results demonstrated that ING4 inhibited estrogen-independent ER activity, suggesting that ING4-low breast tumors recur faster due to estrogen-independent ER activity that renders tamoxifen less effective. This study puts forth fulvestrant as a proposed therapy choice for patients with ING4-low ER+ breast tumors.

PRODUCTO CENTRO AMERICANO: MADE IN HONDURAS By Alma Leiva Master of Fine Arts A thesis submitted in partial fulfillment of the requirements for the degree of Master of Fine Arts at Virginia Commonwealth University. Virginia Commonwealth University, 2011. Thesis Director: Robert Paris, Professor Kinetic Imaging / Photography and Film I was born in 1973 in Honduras, a country under military regime. In 1982 after 20 years of military rule, Honduras finally had democratic elections. During that decade, and as a consequence of the cold war, the kidnapping, torturing, murder and disappearance of civilians became common practices among the Honduran military. Peasant activists, university leaders, union workers and intellectuals were among its favorite targets. In Producto Centroamericano: Made in Honduras I present the viewer with a "product" entrenched in Honduran history; the disappearance of almost two hundred civilians in the 1980's for political reasons. The work also presents the viewer with the imminent threat of the return of this practice after its military coup in 2009. By juxtaposing references of torturing tools such as knives, metal poles, chains and meat hooks, against more frail materials such as paper, wax and fabric, I make allusions to the vulnerability of the individual against such repressive forces. Through the elements presented in the installation, I try to take the viewer on a journey that will hopefully, confront one with one’s own humanity and ultimately with one’s own mortality.

Macrophages are important effector cells of the innate and adaptive immune response and exert a wide variety of immunological functions which necessitates a high level of plasticity on the chromatin level. In response to pathogen-associated molecular patterns (PAMPs) or inflammatory signals macrophages undergo a process of cellular activation which is associated with morphologic, functional and biochemical changes. Toll-like receptors (TLR) are able to sense many different PAMPs. TLR4 is an important sensor for lipopolysaccharide (LPS) which elicits a major portion of the host’s inflammatory response through the activation of many different signaling pathways such as the NF-κB and the MAPK protein kinase pathways RASRAF- MEK-ERK, p38 and JNK. Polycomb group (PcG) proteins are well known chromatin modifiers which function in large complexes and are required to maintain chromatin structure in a transcriptionally repressed state. It has previously been shown that the PcG protein Bmi1 is phosphorylated by 3pK, a downstream effector kinase of the MAPK protein kinase pathways RAS-RAF-MEK-ERK, p38 and JNK. In this work I analyzed the role of Bmi1 as a downstream effector of MAPK signaling during macrophage activation. Unexpectedly a rapid up-regulation on the Bmi1 protein level was observed in bone marrow derived macrophages (BMDMs) after LPS treatment. The Bmi1 induction was associated with transient protein phosphorylation that occured downstream of MAPK signaling. LPS treatment of BMDMs in the absence of Bmi1 resulted in a pronounced increase of IL-10 secretion. This secretion of the anti-inflammatory cytokine IL-10 was associated with increased IL-10 mRNA levels. Furthermore, siRNA mediated knock down of Bmi1 in J774A.1 macrophages also resulted in elevated IL-10 mRNA levels in response to LPS. ChIP analysis revealed that Bmi1 binds to throughout the il-10 locus. Alternative activation of wild type BMDMs via concomitant TLR4 and FcγR activation which triggers high IL-10 expression is paralleled by an attenuated Bmi1 protein expression. These results identify Bmi1 as a repressor of IL-10 expression during activation of macrophages. / Makrophagen sind wichtige Effektorzellen der angeborenen und adaptiven Immunantwort und üben eine große Fülle von immunologischen Funktionen aus. Deshalb benötigen sie eine hohe Plastizität auf Chromatinebene. Als Antwort auf pathogen-assoziierte molekulare Muster (PAMPs) oder andere inflammatorische Signale machen Makrophagen einen zellulären Aktivierungsprozess durch, der mit morphologischen, funktionellen und biochemischen Veränderungen assoziiert ist. Toll-like Rezeptoren (TLR) sind fähig, solche PAMPs zu erkennen. Der TLR4 ist ein wichtiger Sensor für Lipopolysaccharid (LPS). LPS löst eine inflammatorische Antwort des Wirtes durch die Aktivierung vieler verschiedener Signalwege wie zum Beispiel des NF-κB Signalwegs und der MAPK Proteinkinasensignalwege RAS-RAFMEK- ERK, p38 und JNK aus. Polycomb Gruppen (PcG) Proteine arbeiten in großen Proteinkomplexen zusammen und werden benötigt, um das Chromatin in einem transkriptionell reprimierten Zustand beizubehalten. Es konnte gezeigt werden, dass das PcG Protein Bmi1 von 3pK, einer Effektorkinase der MAPK Proteinkinasensignalwege RAS-RAF-MEK-ERK, p38 und JNK, phosphoryliert wird. In meiner Doktorabeit analysierte ich die Rolle von Bmi1 als downstream-Effektor der MAPKSignaltransduktion während der Makrophagenaktivierung. Es wurde unerwarteterweise eine schnelle Induktion von Bmi1 auf Proteinebene in Knochenmarks-Makrophagen (BMDMs) beobachtet. Diese Induktion war MAPK-abhängig und mit einer transienten Proteinphosphorylierung assoziiert. Die LPS-Behandlung der BMDMs in Abwesenheit von Bmi1 hatte erhöhte IL-10 Sekretion zur Folge. Diese erhöhte Sekretion des antiinflammatorischen Zytokines IL-10 war mit erhöhten IL-10 mRNA Expression verbunden. Des Weiteren hatte ein Bmi1 siRNA-vermittelter Gen-Knockdown in J774A.1 Makrophagen eine erhöhte IL-10 mRNA-Expression zur Folge. ChIP Analysen haben gezeigt, dass Bmi1 am ganzen il-10-Locus verteilt binden kann. Eine TLR4 und FcγR vermittelte alternative Aktivierung von BMDMs, die mit hoher IL-10 Expression verbunden ist, führte zu einer attenuierten Bmi1 Proteinexpression. Diese Ergebnisse zusammengenommen weisen Bmi1 als Repressor von IL-10 während der Makrophagenaktivierung aus.

Made available in DSpace on 2016-04-27T19:30:58Z (GMT). No. of bitstreams: 1 Jose do Espirito Santo Dias Junior.pdf: 7092852 bytes, checksum: ece7f9025be578aab2c93f655bf12bfa (MD5) Previous issue date: 2014-08-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This paper deals with the representations bohemian in Belem between 1950-1980, from the understanding of the discourses produced by Para press , cinema , history and memory of their regulars : musicians , bohemians , artists , prostitutes and other social agents involved with these parties . Seek to understand the importance of these houses on the social scene and playfulness of Bethlehem during the chronological cut above, considering the geographical changes of the city and the symbolic and cultural representations created about these establishments. Thus , we intend to understand the history of men and women who made these spaces , its locus par excellence of entertainment and leisure , the sacred place of amusement weekend and early mornings , a universe marked by bohemian social relations of pleasure , joy and entertainment that , was the victim of repressive and vigilant eyes of the powers that be, always with the justification defense of "good manners".." A cultural environment wrapped in beverages, boleros, meringues, bohemians , whores and bums , who helped achieve the specter of danger and adventure, but at the same time , built romantic and nostalgic images of these places / O presente texto trata das representações boêmias em Belém do Pará entre 1950-1980, a partir da compreensão dos discursos produzidos pela imprensa paraense, pelo cinema, pela história e memória de seus frequentadores: músicos, boêmios, artistas, meretrizes e demais agentes sociais envolvidos com essas festas. Busco entender a importância dessas casas no cenário social e lúdico de Belém durante o recorte cronológico supracitado, considerando as transformações geográficas da cidade e as representações simbólicas e culturais criadas acerca desses estabelecimentos. Assim, pretendo compreender a história de homens e mulheres que faziam desses espaços, seu lócus por excelência de entretenimento e lazer, o lugar sagrado das diversões de final de semana e das madrugadas, um universo boêmio marcado por relações sociais de prazer, alegria e entretenimento, que, não raras vezes, foi vítima dos olhos vigilantes e repressores dos poderes instituídos, sempre com a justificativa de defesa dos bons costumes . Um ambiente cultural envolto em bebidas, boleros, merengues, boêmios, meretrizes e festeiros, que ajudaram a lograr o espectro da periculosidade e aventura, mas que, ao mesmo tempo, construíram imagens nostálgicas e românticas desses lugares

The phenomenon of cell type-specific DNA methylation has received much attention in recent years and a number of DNA methylation differences have been described between cells of the immune system. Of particular interest when studying DNA methylation are CpG islands (CGIs) which are distinct from the rest of the genome due to their elevated CpG content, generally unmethylated state and promoter association. In the instances when they become methylated this is associated with gene repression although it is unclear the extent to which differential methylation corresponds to differential gene expression. I have used an immune system model to assess the role of CGI methylation and the role of the methylation reader MBD2 in regulation of gene expression. A relatively small number of DNA methylation differences were seen between immune cell types with the most developmentally related cells showing the fewest methylation differences. Interestingly, the vast majority of CGI-associated cellspecific methylation occurred at intragenic CGIs located, not at transcription start sites, but in the gene body. Increased intragenic CGI methylation tended to associate with gene repression, although the precise reason for this remains unclear. Most differentially methylated CGIs were depleted for the active chromatin mark H3K4me3 regardless of their methylation state but some of these were associated with the silencing mark H3K27me3 when unmethylated. These findings suggest that intragenic CGIs are a distinct class of genomic element particularly susceptible to cell type-specific methylation. I also looked at the effect of removing the methyl- CpG binding domain protein MBD2 from immune system cells. Immune cells from Mbd2-/- mice showed a number of previously uncharacterised phenotypes as well as a number of differences in gene expression compared to wild-type animals. Most of these genes increased their expression in the absence of MBD2 consistent with MBD2’s role as a transcriptional repressor and Mbd2-/- Th1 cells showed increases in histone H3 acetylation compared to wild-type Th1 cells. This work provides an insight into the role played by cell-specific CGI methylation and MBD2 in regulating gene expression.

A pesquisa apresenta a teoria marcuseana do poder como perspectiva crítica no debate contemporâneo acerca do conceito foucaultiano de biopolítica. À primeira vista, tal relação parece controversa, ao reconhecer que Foucault desenvolve seu conceito paralelamente à crítica contra Marcuse. Ora, o conceito foucaultiano de biopolítica descreve jogos de poder como administração dos corpos e também como um modo de cálculo da vida da população. Tal concepção contraria diretamente a hipótese marcuseana do poder repressivo, um modelo crítico que tem em vista uma camada verdadeira e subjacente de poder, recalcada nas formações sociais e subjetivas estabelecidas. Com esse quadro, como reunir os dois autores na crítica do poder, assumindo a biopolítica como premissa da teoria do poder? De fato, a aproximação seria impossível ao partir da aposta marcuseana em uma civilização nãorepressiva, presente em Eros e Civilização. Contudo, com a análise do avanço da racionalidade instrumental no pós-Guerra em O Homem Unidimensional, Marcuse avalia a possibilidade de um poder não-repressiva. Afinal, na nova ordem social não se apresenta mais um controle repressivo dos corpos, mas sim uma excitação da vida e dos corpos em movimento. Seria este um sinal de concordância entre os autores? E ainda, dada esta nova correspondência, é possível aproveitar a crítica foucaultiana à biopolítica para redimensionar não apenas as reflexões de Marcuse sobre o poder, como também as passagens contemporâneas entre a teoria crítica e a genealogia do poder? / This research presents Marcuses theory of power as an essential perspective for the contemporary debate on the Foucaultian concept of biopolitics. This would seem a rather controversial choice at a first glance, as, admittedly, the development of Foucaults concept occurred alongside that of his critique of Marcuse. Indeed, Foucaults conception of biopolitics describes games of power that include the administration of the bodies and the calculated management of the life of a given population a notion entirely adverse to Marcuses repressive hypothesis of power, a critical model that assumes a real, subjacent layer of power that is repressed in established social and subjective formations. Given these differences, as well as an adoption of biopolitics as a fundamental premise for a theory of power, how are the two authors to be brought together for a critique of power? Such an approximation would certainly be impossible in light of Marcuses arguments, in Eros and Civilization, for the possibility of a non-repressive civilization. Still, through the analysis of the advancement of instrumental rationality in the postwar period conducted in One- Dimensional Man, Marcuse will revise his former perspective on non-repressive power; after all, the new social order no longer features a repressive control of bodies, but rather an excitation of life, and of bodies to motion. Could that be understood as a sign of agreement between the authors? Furthermore, given this new correspondence, would it be possible to employ Foucaults critique to add dimensions not only to Marcuses reflections on power, but to contemporary mediations between critical theory and the genealogy of power?

A la faveur d'un accroissement de la répression et de l'introduction des techniques de managérialisation (telles que la comparution sur reconnaissance préalable de culpabilité) au sein de l'institution judiciaire, les magistrats des juridictions lyonnaise, roannaise et stéphanoise adoptent des pratiques standardisées en matière de répression de la délinquance routière. Cette apparition d'un modèle managérial et standardisé du traitement de la délinquance routière, privilégiant une réponse pénale systématique, rapide, normée et sévère, redéfinit le cadre de travail de tous les acteurs de la chaîne pénale, en l'occurrence des forces de l'ordre, des parquetiers, des juges et des avocats. Plus précisément, remettant en cause les principes d'individualisation de la peine et d'autonomie propre à la profession de magistrat, les parquetiers et, par effet domino, les juges alignent leurs pratiques sur les prescriptions ministérielles qui sont de plus en plus répressives. Ce travail montre que la déclinaison de la politique pénale en matière de délinquance routière au niveau local est le produit d'une action top-down, traduisant un «Etat en action » et non pas en « interaction ». La politique pénale de sécurité routière et le traitement de la délinquance routière restent plus que jamais des secteurs régaliens dans lesquels l'Etat veut réaffirmer son leadership. / As a consequence of the increase in repression and of the implementation of management standards(such as the plea bargaining) in Justice, the substitutes of the prosecutor and the judges adopt standardized practices as regards repression of the road delinquency in Lyon, Roanne and Saint-Etienne. The implementation of management standards as regards the road delinquency, privileging systematic and fast penal answer, redefines the framework of ali the actors of the criminal justice system (police, substitutes of the prosecutor, judges and lawyers). More precise! y, the substitutes of the prosecutor and the judges put into question fundamental principles such as individualized judgments and autonomy. They align their practices on the ministerial directives, which are increasingly repressive. This work shows that the variation in criminal policy as regards road delinquency at the local leve! is the product of a top-down action, translating a "State into action" and not in "interaction". The cri minai policy of road safety and the treatment of road delinquency remain sectors in which the State wants to reaffirm its leadership.

Upon oncogenic stress, the tumor suppressor Arf can induce irreversible cell cycle arrest or apoptosis, depending on the oncogenic insult. In this study, it could be shown that Arf interacts with Myc and the Myc-associated zinc-finger protein Miz1 to facilitate repression of genes involved in cell adhesion. Formation of a DNA-binding Arf/Myc/Miz1 complex disrupts interaction of Miz1 with its coactivator nucleophosmin and induces local heterochromatinisation, causing cells to lose attachment and undergo anoikis. The assembly of the complex relies on Myc, which might explain why high Myc levels trigger apoptosis and not cell cycle arrest in the Arf response. This mechanism could play an important role in eliminating cells harboring an oncogenic mutation. Arf furthermore induces sumoylation of Miz1 at a specific lysine by repressing the desumoylating enzyme Senp3. A sumoylation-deficient mutant of Miz1 however does not show phenotypic differences under the chosen experimental conditions. Myc can also be modified by Sumo by multisumoylation at many different lysines, which is unaffected by Arf. The exact mechanism and effect of this modification however stays unsolved. / Der Tumorsuppressor Arf wird durch onkogenen Stress induziert und kann entweder einen irreversiblen Zellzyklusarrest oder Apoptose auslösen. In dieser Arbeit konnte gezeigt werden, dass Arf mit Myc und dem Myc-interagierenden Zinkfingerprotein Miz1 assoziiert und dadurch Gene der Zelladhäsion reprimiert. Die Ausbildung eines DNA-bindenden Arf/Myc/Miz1 Komplexes verhindert eine Interaktion von Miz1 mit seinem Koaktivator Nucleophosmin und führt zur lokalen Ausbildung von Heterochromatin, was zum Ablösen der Zellen und schließlich zur Anoikis führt. Die Komplexbildung setzt die Beteiligung von Myc voraus, was erklären könnte warum hohe Mengen an Myc über Arf Apoptose und nicht Zellzyklusarrest auslösen. Dieser Mechanismus könnte eine wichtige Rolle bei der Eliminierung von Zellen mit einer onkogenen Mutation spielen. Arf induziert darüber hinaus die Sumoylierung von Miz1 an einem bestimmten Lysin indem es das desumoylierende Enzyme Senp3 inhibiert. Eine Mutante von Miz1 die nicht mehr sumoyliert werden kann zeigt jedoch in den durchgeführten Untersuchungen keinen anderen Phänotyp als Wildtyp Miz1. Myc kann ebenfalls an vielen verschiedenen Lysinen mit Sumo modifiziert werden, wobei Arf jedoch keine Rolle spielt. Der genaue Mechanismus und Effekt dieser Modifikation konnte jedoch nicht geklärt werden.