Identification of genes that rescue Dbp5-Nup159 temperature-sensitivity
Sciences and Mathematics, College of
Biology, Department of
SURS Faculty Advisor
Dr. Rebecca Adams
The export of mature mRNA through the nuclear pore complex (NPC), as directed by the mRNA export protein Mex67, is necessary for successful protein synthesis of the transcript in the cytoplasm. In S. cerevisiae, a protein called Dbp5 is responsible for removing Mex67 from the mRNA after it passes through the NPC and into the cytoplasm, providing directionality to the export process. Dbp5 is localized at the cytoplasmic side of the NPC through interaction with the NPC protein Nup159. The exiting RNA comes into contact with Dbp5 right after export for removal of Mex67, permitting its removal once the transcript reaches its destination. Dbp5 can also be found inside the nucleus and throughout the cytoplasm, where it has been implicated in the processes of transcription and translation, respectively. However, the molecular details of Dbp5’s function in these subcellular locales is unknown. The goal of this study was to identify other functions of Dbp5 in these contexts. I hypothesized that Dbp5 functions to remove other proteins from RNA in these other cellular compartments. In order to uncover these functions, I have followed up on a previously- performed multicopy suppression screen with a temperature-sensitive Dbp5-Nup159 fusion strain in which Dbp5 is anchored at the NPC. Our specific goal was to narrow down the rescuing gene from the plasmid that encodes several genes. To do this, I used a series of restriction enzymes to cut genes out of the plasmid. The resulting plasmids will then be re-transformed into the Dbp5-Nup159 strain to assess rescue of temperature-sensitivity. We anticipate that the rescuing gene will aid in identification of nuclear and cytoplasmic protein targets of Dbp5.
Harbison, Zoe and Adams, Rebecca, "Identification of genes that rescue Dbp5-Nup159 temperature-sensitivity" (2022). Science University Research Symposium (SURS). 24.