I. Synthesizing and Identifying Small Molecule Probes for Targeting Transcriptional Co-factors II. Design and Implementation of a Peer-Led Practical Research Ethics Module for Teaching Graduate Research Ethics.

Abstract

Activated transcription is a complex process that is governed by the interactions of several different proteins. As misregulation of transcription has been observed in many diseases, understanding the roles of the proteins involved in transcription will aid in the design of therapeutics that target aberrant gene expression. One protein-protein interaction that is essential in governing transcription is that of a DNA bound transcriptional activator with co-activator proteins. Two co-activators, CBP and p300, are highly homologous proteins that play key roles in several cellular processes including cell growth, differentiation, cell-cycle regulation and apoptosis. Emerging evidence suggests that these proteins also play a role in the maintenance of pluripotency and self-renewal in embryonic and hematopoietic stem cells. Therefore, there is much interest in identifying small molecules probes that can perturb the functions of CBP and p300 in defined ways.

As CBP/p300 have been known to participate in a wide variety of binding interactions, it is unlikely that a single molecular scaffold will be sufficient to perturb all of these interactions. The use of building block substrates that can be further taken on to form a variety of structures should be employed in the synthesis of small molecule libraries that can be screened for their ability to modulate transcription. A novel methodology for the synthesis of one such building block substrate, an allenamide, is reported here. Additionally, small molecule modulators of transcription are employed in examining the roles of CBP and p300 in maintaining pluripotency and self-renewal in cancer initiating cells (CICs). Studies carried out with these probes, along with RNAi directed against CBP or p300, demonstrate that p300 is required for Nanog-driven gene expression and that disrupting the function of p300 and/or CBP affects the self-renewal capabilities of CICs. Finally, this work takes a step back from transcription to address an issue faced by all scientists: research ethics. The design and implementation of a peer-led module for first year graduate students on practical research ethics is discussed. Student responses during group discussions suggest that this method is a successful strategy for teaching research ethics to graduate students.