Sciences and Mathematics, College of
Biology, Department of
BURS Faculty Advisor
During an mRNA’s lifecycle, RNA-binding proteins (RBP) are added and removed in order to facilitate export from the nucleus to the cytoplasm through the nuclear pore complex (NPC). During the RNA export process, a class of proteins termed Dead-box proteins (Dbps) are strategically located at the cytoplasmic face of the NPC to remove mRNPs from the mRNA-protein complex, providing one way directionality for export in the process termed mRNA-protein (mRNP) remodeling. Specifically, Mex67 is an RBP which ferries the transcript through the nuclear pore complex and is removed by Dbp5, which is located at the cytoplasmic side of the NPC. Dbp5 is localized here through interactions with nup159 which is an integral NPC protein. Although Dbp5’s function in the process requires NPC localization, Dbp5 is also dispersed throughout the nucleus and the cytoplasm. Previous research has found that through deleting wild type Dbp5 from the S. cerevisiae genome, and generating a Dbp5-nup159 fusion strain, the cell loses its nuclear/ cellular dbp5’s function which causes the cell to have a temperature-sensitive growth defect. We hypothesize that this lost localization results in a growth defect because Dbp5 serves to remove other RBPs from mRNA at other phases of its life cycle. To identify this putative RBP, previous students have performed a multi-copy suppression screen where it was anticipated that over expression of the RBP will restore growth at higher temperatures. This screen resulted a plasmid that rescued temperature-sensitivity in the Dbp5-nup159 fusion strain, however the plasmid contains many genes, and the specific rescuing gene has not been identified. By digesting the plasmid with select restriction enzymes to systematically remove regions of the rescuing plasmid, we have generated a series of truncated plasmids. We anticipate that these plasmids will help identify the gene responsible for rescuing the growth defect.
Hooks, Jonah and Adams, Rebecca, "Uncovering Genes Involved in Non-NPC-Associated Dbp5 Function" (2023). Belmont University Research Symposium (BURS). 235.