Understanding Iron Regulation Through Expression Vectors In Yeast Cells

Iron regulation is extremely vital to human health and cellular functions. Excess iron can cause diseases such as hemochromatosis, while iron deficiency can lead to anemia. Almost 50% of the worldwide population suffers from iron regulatory difficulties. The main purpose behind this project involves finding ways to reduce that percentage. In this specific study, iron regulation is examined by characterizing different genes and transcription factors that affect iron homeostasis in yeast cells. The goal of this project is to clone Aft1, a transcription factor that turns on iron uptake genes when iron levels are low. Once activated, the gene is inserted into a yeast overexpression plasmid with a myc epitope tag using the Gibson assembly approach. Upon completion, this new plasmid will allow the overexpression of Aft1 from its native cell with a tag that allows easy purification. Previous results indicate Aft1 works with Yap5, which turns on iron sequestration genes when iron levels are high to prevent iron overload. By cloning these genes and purifying these proteins for biochemical analysis, further research can be done to characterize the molecular mechanisms that govern Aft1 and Yap5 activity in cells. This information will provide insight into the basic biology of iron metabolism, which is useful for developing breakthroughs in solving these regulatory issues