Poising of RNA Polymerase II: Features and Functions

Colorado State University, Fort Collins CO

Investigators

Abstract

Intellectual Merit. The initial discovery of the occupancy of RNA polymerase II (RNAPII) at certain genes prior to their transcriptional activation occurred over a quarter century ago in Drosophila. Due to recent advances in whole genome analyses, occupancy of RNAPII in an inactive state can be detected in organisms across the evolutionary spectrum and at diverse sets of genes. The goal of this project is to investigate post-recruitment mechanisms of gene regulation by focusing on poised RNAPII in yeast and the response to oxidative stress as the regulatory system. These studies take full advantage of the available approaches (genetics, molecular biology, biochemistry, cell biology, genomics), and are amenable to researchers at a variety of levels (graduate, undergraduate and elementary school students). Thus, the transcriptional response to oxidative stress will be investigated with a focus on the pre-bound and poised RNAPII complex by testing two overarching hypotheses: 1) Poised promoters share common features that are distinct from recruitment-regulated promoters; and 2) Poised promoters offer functional advantage(s) for the regulation of gene expression. Since post-recruitment regulation is widespread in humans and other organisms, these findings will significantly impact the field of gene expression with exceptional potential to alter our overall views. Thus, these studies are extremely timely and highly significant. Broader Impacts. The project has broad impact in at least four ways. First, the studies offer suitable projects for undergraduate trainees, who may have time constraints as well as limited laboratory experience. Second, studies with higher technical and time demands provide outstanding cross-training experiences for graduate students in a number of different disciplines (genetics, genomics, cell and molecular biology). These techniques are highly versatile. Trainees will utilize these techniques in the future if they choose to investigate other critical genetic mechanisms (DNA repair, replication, recombination, RNA metabolism, epigenetics, etc.). Third, to propagate the tradition of outreach, undergraduate and graduate students will be involved in taking gene expression studies into elementary school classrooms. As part of a previously funded NSF project, fifth graders and their teachers helped to develop a novel outreach program called 'Biochemistry is Elementary'. This innovative program has eight separate 1 hour sessions involving hands-on activities designed to explore basic concepts of scientific reasoning involving biochemistry and yeast genetics. Due to the inexpensive and biologically safe reagents and the detailed workbooks created during the project, many scientists could offer something similar to their local community if they received training on the program. As such, this program will be expanded to include an interactive website. Finally, results from this project will be published in broad based scientific journals, and presented at local and international meetings. Taken together, this project directly supports the mission of the NSF to advance discovery and understanding, while promoting teaching, training and learning.