Elucidation of the interactions between the transcription factor E2A and the transcriptional co-activator CBP/p300

E2A is a transcription factor that plays a particularly critical role in lymphopoiesis. The chromosomal translocation 1;19, disrupts the E2A gene and results in the expression of the fusion oncoprotein E2A-PBX1, which is implicated in acute lymphoblastic leukemia. Both E2A and E2A-PBX1 contain two activation domains, AD1 and AD2, which comprise conserved ΦxxΦΦ motifs where Φ denotes a hydrophobic amino acid. These domains function to recruit transcriptional co-activators and repressors, including the histone acetyl transferase CREB binding protein (CBP) and its paralog p300. The PCET motif within E2A AD1 interacts with the KIX domain of CBP/p300, the disruption of which abrogates the transcriptional activation by E2A and the transformative properties of E2A-PBX1. The generation of a peptide-based inhibitor targeting the PCET:KIX interaction would serve useful in further assessing the role of E2A and E2A-PBX1 in lymphopoiesis and leukemogenesis. An interaction between E2A AD2 and the KIX domain has also been recently identified, and the TAZ domains of CBP/p300 have been shown to interact with several transcription factors that contain ΦxxΦΦ motifs. Thus the design of an inhibitor of the E2A:CBP/p300 interaction requires the full complement of interactions between E2A and the various domains of CBP/p300 to be elucidated. Here, we have used nuclear magnetic resonance (NMR) spectroscopy to determine that AD2 interacts with KIX at the same site as PCET, which indicates that the E2A:KIX interaction can be disrupted by targeting a single binding site. Using an iterative synthetic peptide microarray approach, a peptide with the sequence DKELQDLLDFSLQY was derived from PCET to interact with KIX with higher affinity than the wild type sequence. This peptide now serves as a lead molecule for further development as an inhibitor of the E2A:CBP/p300 interaction. Fluorescence anisotropy, peptide microarray technology, and isothermal titration calorimetry were employed to characterize interactions between both TAZ domains of CBP/p300 and the PCET motif and AD2 of E2A. Alanine substitution of residues within PCET demonstrated that the ΦxxΦΦ motif is a key mediator of these interactions, analogous to the PCET:KIX interaction. These findings now inform future work to establish possible physiological roles for the E2A:TAZ1 and E2A:TAZ2 interactions.

CERAMICS IN BRITAIN (1840-90): MEANINGS AND METAPHORS

The intention of this thesis, “Ceramics in Britain (1840–90): Meanings and Metaphors” is to present new approaches for interpreting ceramics in nineteenth-century Britain by situating, problematizing, and contextualizing pottery and porcelain in the popular debates of the day within the methodologies of material culture, design, cultural and art histories. I ask how did ceramics—portable, functional, and often decorative objects—contribute to shaping modes of experiences? Crockery, tableware and blue-white-porcelain, admittedly largely mediated in texts and paintings, are at the centre of this research to examine how they imposed symbolism and influenced the engagement of their subjects beyond their intended meanings and functions. This thesis tracks a common rhetoric shared by writers and artists across genres and understood by readers and viewers: crockery in the cupboard, on the mantel, the table or the floor were popular motifs exemplifying class, gender, character, etiquette, and taste. This thesis also seeks to map ceramics’ relations with other objects and people depicted. Their meanings and metaphors changed, depending on their exchange with other objects in the room and who uses them. The conventions of representing ceramics dictated a particular grammar that writers and artists used, critiqued, discarded or personalized. The examination of ceramics mediated in text and image especially in comparison with extant objects invites a deeper probing of both material culture and artistic practice, which helps to situate the agency of the ceramic objects themselves. Also this thesis, in attempt to explore new methodological approaches for ceramic studies, examines the social life of the mid-Victorian relief-moulded “Minster” Jug in the Gardiner Museum in Toronto. The product originating in Staffordshire in 1843 and exported to the colonies holds significance due to its multiple life histories. Viewing the “Minster” through the lenses of curator, collector, consumer, and critic its layered lives unfold to reveal the protocols of museum praxis as well as important aspects of mid-nineteenth-century British society related to design reform, gender, imperialism and consumption patterns. This thesis contends that the British experienced ceramics in sometimes unexpected ways, unrelated to their original purpose, such as tools of violence or containers of solace, and transformative fantasy.

WAKING (TO) THINKING BEING: A STUDY OF EDUCATION, METAPHYSICS, MIND, AND LANGUAGE

This dissertation articulates the basic aims and achievements of education. It recognizes language as central to thinking, and philosophy and education as belonging profoundly to one another. The first step is to show that although philosophy can no longer claim to dictate the foundations of knowledge or of disciplines of inquiry, it still offers an exceptionally general level of self-understanding. Education is equally general and faces a similar crisis of self-identity, of coming to terms with reality. Language is the medium of thought and the repository of historical mind; so a child’s acquisition of language is her acquisition of rational freedom. This marks a metaphysical change: no longer merely an animal, she comes to exercise her powers of rationality, transcending her environment by seeking and expressing reasons for thinking and doing. She can think about herself in relation to the universe, hence philosophize and educate others in turn. The discussion then turns to the historical nature of language. The thinking already embedded in language always anticipates further questioning. Etymology serves as a model for philosophical understanding, and demonstrates how philosophy can continue to yield insights that are fundamental, but not foundational, to human life. The etymologies of some basic educational concepts disclose education as a leading out and into the midst of Being. The philosophical approach developed in previous chapters applies to the very idea of an educational aim. Discussion concerning the substantiality of educational ideals results in an impasse: one side recommends an open-­ended understanding of education’s aims; the other insists on a definitive account. However, educational ideals exhibit a conceptual duality: the fundamental achievements of education, such as rational freedom, are real; but how we should understand them remains an open question. The penultimate chapter investigates philosophical thinking as the fulfillment of rational freedom, whose creative insights can profoundly transform our everyday activities. That this transformative self-understanding is without end suggests the basic aims of education are unheimlich. The dissertation concludes with speculative reflection on the shape and nature of language, and with the suggestion that through education reality awakens to itself.