Post-transcriptional Control of Immunoglobulin Expression

University Of Kentucky Research Foundation, Lexington KY

Investigators

Abstract

Intellectual Merit: The long-term goal of this research is to understand the regulation of gene expression, focusing specifically on the connections between gene transcription and RNA processing. While a substantial base of knowledge has been accumulated about these processes individually, how they are coupled and how this coupling affects regulation has yet to be determined. This project will explore the mechanistic interconnections between RNA processing events (splicing and cleavage-polyadenylation) and transcription elongation and termination in the immunoglobulin M (IgM or µ) gene. The µ gene is an excellent model system in which to address these questions since its expression is regulated during B lymphocyte maturation at multiple steps, including RNA splicing, cleavage-polyadenylation and transcription termination. An extensive collection of well-characterized modified µ genes, multiple reagents to analyze µ gene expression and microarray data from B cells before and after being stimulated to differentiate, are available in the PI's lab for this work. The project will identify trans-regulators of µ alternative RNA processing based on microarray data and gene expression experiments, determine how RNA processing signals affect RNA polymerase II elongation, using a combination of nuclear run-on and high-resolution chromatin immunoprecipitation (ChIP) assays and characterize changes in factors associated with elongating pol II over wild-type and modified µ genes in B cells and plasma cells by high-resolution ChIP experiments. These studies will lead to a better understanding of the connections between transcription and RNA processing in the µ gene and will shed new light on these connections, which also regulate the expression of many other genes. Broader Impacts The broader impacts of this research include the involvement of graduate, undergraduate, and possibly high school students in the research, as well as their participation in local and national scientific meetings. The majority of students associated with the PI's lab over the past eighteen years have been female. An on-going collaboration with a former graduate student from this lab, who is currently an Associate Professor at Middle TN State University, will continue; the microarray data has been the basis for projects in undergraduate lab courses and individual student independent research projects. The PI of this grant continues to support science in the broader community by participating as a science fair judge and visiting high school classrooms. In addition, as a part-time Associate Vice President for Research at the University of Kentucky and member of the KY Academy of Sciences, the PI supports research in institutions across the state of KY. The data and resources generated from this project will be shared widely with the scientific community through national and international meeting presentations and publications.