Molecular interactions between Parkin and FBXO7 and links with induced dopaminergic neuron degeneration in Parkinson's disease.
Tang, Alyssa Meiyan.
Date of Issue2013
School of Biological Sciences
Mitophagy has received much attention regarding the pathogenesis of PD. However, mitophagy studies have been encumbered by cell quantities, which will interfere with mitochondria readings. By double-staining cells with NAO and Hoechst after inducing mitophagy with FCCP, we developed a simple but accurate method for studying mitophagy, which idealistically, could be a new standard for studying mitophagy. The FCCP treatment concentration and time was optimized to minimize cell death and induce mild levels of mitophagy. Using the conditions optimized we found that FBXO7, a protein not previously linked to mitophagy, plays a role in promoting mitophagy after treating with low concentrations of FCCP. R498X, a mutant form of FBXO7, lost this ability to control mitochondria quality. Finally we conducted a preliminary study on the effect of co-transfecting WT Parkin and FBXO7 with mutant FBXO7 and Parkin to see if these proteins can rescue each other’s mitophagy functions. In conclusion, in addition to discovering that FBXO7 can induce mitophagy, we have established a platform for further investigation on how Parkin and FBXO7 WT and mutant proteins interact and influence mitophagy. This will shed light on the function of FBXO7, its relationship with parkin, and the aetiology of PD.
Final Year Project (FYP)
Nanyang Technological University