Mechanism and Regulation of Core Promoter Recognition and Promoter Escape during Transcription by RNA Polymerase II

University Of Colorado At Boulder, Boulder CO

Investigators

Abstract

Intellectual merit: The expression of genes that are encoded by DNA is essential for sustained life. The first step in gene expression is making an RNA copy of the gene through a process known as transcription. During this process, the enzyme RNA polymerase II, using DNA as a template, synthesizes mRNA molecules. The broad goal of this research is to arrive at a detailed understanding of the factors and the mechanism of the transcription reaction and how they can be controlled. Such an understanding is important because transcription is a principal control point for regulating gene expression during growth, development, and normal cellular metabolism. Specifically, this research will identify and characterize the function of a factor in human nuclei that can accelerate the rate of the slow step in transcription, hence defining a new mechanism of regulation. This research will also investigate how large complexes containing proteins and DNA, which are required for transcription to occur, assemble at genes which are poised to be transcribed. The methods used in these studies center on a well-established human transcription system that is assembled in vitro from highly purified proteins. The experiments utilize detailed quantitative assays specifically developed to study the early steps of transcription and the assembly of proteins on DNA. The research will impact the understanding of how protein-DNA complexes assemble at genes, and how the early steps in RNA synthesis might be regulated in cells. The knowledge derived from this project has the potential to stimulate new studies of transcriptional regulation in complex biological systems. Broader impacts: In addition to the research described above, the project will involve development of an experimental module for an undergraduate biochemistry laboratory. This module will provide students with experience in designing and implementing a hypothesis-driven experiment investigating protein-DNA interactions. Moreover, students will be exposed to new experimental techniques, thereby enhancing the scientific infrastructure of a teaching laboratory course. Developing an experimental module for an undergraduate laboratory underscores the importance of integrating research and education in order to promote teaching, training, and learning. In addition, implementation of the research plan will provide opportunities to devote a significant amount of time to the training of graduate students in experimental design, interpretation of data, creative thought, preparation and writing of manuscripts, and giving formal and informal oral presentations.