The identity, activity, and longevity of plant cells is largely governed by the assortment of proteins they contain. The synthesis of proteins is a highly orchestrated process that begins in the cell nucleus with gene transcription and continues in the cytoplasm following messenger RNA (mRNA) export from the nucleus. In the cytoplasm, an mRNA may be stable or unstable, and may be sequestered or actively translated by ribosomes into protein. The stability or translation of mRNAs in multicellular plants also depends on developmental or environmental signals. The goal of this project is to clarify how this cytoplasmic post-transcriptional regulation occurs, and specifically to analyze the roles of RNA binding proteins (RBPs) in these processes. In this Arabidopsis 2010 project, three research groups will collaborate to characterize the specific roles of more than 40 RBPs. A major focus is to identify the mRNAs that are bound by specific a RBP in an RNA-protein (RNP) complex, and to identify other proteins within the RNP complex. In addition, select RBPs will be analyzed for their roles in developmental networks and in dynamic responses to changes in cellular status due to environmental stress. The study will include functional analyses of RBPs through use of mutants. Methods will include deep sequencing of the mRNAs associated with specific RBPs and identification of the mRNA sequences recognized by the RBPs. These studies will combine advanced bioinformatic, molecular and genetic approaches.
Broader Impacts The broader impacts from this project fall into three areas. First, the project will engage graduate students and postdoctoral researchers in state-of-the art technologies in molecular and cellular biology as well as bioinformatics. In addition, undergraduate researchers will be mentored in research projects, and scientists and teachers will be provided summer workshops on cellular mechanisms of gene expression. Second, the outcomes of the research will be made available to the larger scientific community through the establishment of a web site (Plant mRNP-Net), which will allow mRNA-protein network mining and include resources for educators. Third, the project will work directly with The Leonardo, a Utah science museum, to assist in development of a classroom demonstration linked to the 9th grade curriculum.
