Please use this identifier to cite or link to this item: http://hdl.handle.net/1946/12145
Temperature sensitive degron strains for Rpt1, Rpt4, Rpt6 and the effect on induced GAL1-10 expression
The 26S proteasome is composed of the 19S and 20S particles. The 20S particle is the protein degrading complex while the 19S particle plays regulatory role. The 19S control complex is composed of 17 subunits including 6 ATPases of the AAA- family. These ATPases, called Rpt1-6, are relatively well conserved between eukaryotes, have similar amino acid sequences and are essential for viability in yeast. Recent studies have suggested that the 19S particle plays a role in the early steps of induced transcription of the GAL gene family. The subject of this study was the role of these ATPases and their effect on induced gene expression of the GAL genes. A heat sensitive degron system was employed to ensure the degradation of the Rpts in a series of experiments where GAL expression was induced after degradation of the Rpts. The results were confirmed with western blots and evaluated with real-time PCR. The final results, while not what was expected based on the literature, turned out to be curious and interesting.