SGER: Ribosomal Frameshifting and Cellular Gene Expression

Rutgers, The State University Of New Jersey-Rbhs-Robert Wood, Piscataway NJ

Investigators

Abstract

Dinman 0084559 The project involves two major paradigm shifts: 1) that certain cellular mRNAs use programmed -1 ribosomal frameshifting (PRF) as a post-transcriptional regulatory mechanism, and 2) that -1 PRF is used to regulate two intracellular signaling pathways (the Jun Kinase and Stress Activated Protein Kinase pathways) that play critical regulatory roles in the control of cell growth, division, differentiation and apoptosis. A bioinformatic strategy combining genome DNA database analysis and DNA microarray technology will be used to test this hypothesis. Computer analyses of the large DNA databases have been used to identify and generate databases of cellular genes that contain potential -1 PRF signals. This project utilizes a DNA microarray approach to identify cellular mRNAs that are specifically stabilized by addition of anisomycin, a drug that specifically inhibits -1 PRF and activates these two signaling pathways. Initial experiments will be done using a yeast-based system due to its experimental malleability. These will allow for testing of the general hypothesis and fine-tuning of the technology. From there, the project will continue using human cell lines. The resulting databases of anisomycin- stabilized mRNAs will be cross-referenced with the database of -1 PRF signal containing- genomic sequences from both yeast and humans. It is envisioned that this approach will identify the mRNAs that serve as the critical checkpoints in the regulation of these essential cellular signaling pathways. Positive results may have significant impact on our understanding of many of the underlying causes of birth defects, cancer and aging.