Our goal is to understand the basic machinery which processes pre-mRNA in the nucleus and apply that understanding towards analysis of how pre-mRNA processing is regulated. Unlike initiation of transcription, which is essentially an on-off type of switch. post-transcriptional control of gene expression allows mammalian cells to produc, from a single transcription event, an enormous variety of protein products. This is because a single pre-mRNA transcript can be alternatively spliced and/or polyadenylated resulting in different combinations of exons being assembled into a mature mRNA. Additional levels of control are also possible since alternative processing of pre-mRNAs can exclude or include RNA sequence elements which differentially regulate mRNA stability, localization and translational efficiency. Alterations and errors of pre-mRNA processing can lead to aberrant gene expression and disease. Both pre-mRNA and mRNA have been the primary targets for therapeutic methods based on antisense RNA and ribozyme technologies. By understanding the mechanisms underlying these processes we will contribute towards improving these therapies.
Because lower, single-cell eukaryotes rarely regulate or alternatively process their pre-mRNAs we focus nearly exclusively on mammalian systems. Methodologies include both biochemical, bioinformatic and reconstitution analysis coupled with studies in tissue culture cells. The goal is to identify both cis-acting RNA sequence elements as well as trans-acting protein factors and understand how they combine to result in regulated pre-mRNA processing. Additionally, we perform extensive biochemical analysis of RNA-protein interactions.
