Dysfunction of Protein Homeostasis: Factors in uencing the Formation of alpha - Synuclein (Lewy Bodies) in Arabidopsis thaliana plants
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BACKGROUND:Protein homeostasis or proteostasis under the conscious control of a network of proteins including chaperones, transporters, ubiquitin-dependent proteasome control the proper expression, folding, translocation and clearance. This is necessary to keep an organism's health and and proteome in the functional state. Therefore, a derangement could possibly lead to aggregation or fibrilization of these proteins which is manifested in most of neuropathies. OBJECTIVES: Our objective was to determine the possible factors that could infl uence the accumulation of α-synuclein in Arabidopsis thaliana plant. METHODS: Three different Arabidopsis thaliana plant (Wild-type(WT) and 2 mutants - ISUSN5 and E46K12) were used for the study. Two sampling methods were used. In sampling method 1, 0.1 g of each of young and old leaves were harvested from the different plants after 4 months. Extracts were prepared from each of them to determine proteasome activity. In sampling method 2, the mutant plants, ISUSN5 and E46K12 were sectioned or divided into 4 quarter. Each quarter was harvested at an interval of 4 weeks within the 4 months period. Each quarter represented the different ages of the plants. Unlike the mutant plants, the whole plant was harvested at the first, second, third and fourth month. RESULTS: The average Relative Fluorescent Unit (RFU) measurement for young were and old leaves for the WT, ISUSN5 and E46K12 were 43568.29, 37182, 96420 respectively for the young leaves and 33982.41, 24145.43 and 159108.15 respectively for sampling method 1. In the second sampling method, the RFU measurements for the WT, ISUSN5 and E46K12 were 17981.05, 35052.42 and 18474.20 respectively. CONCLUSION: RFU measurements were higher in the young leaves than in the old leaves in the WT and ISUSN5 for sampling method 1. In sampling method 2, the average RFU measurement as a function of the proteasome for the first, second, third and fourth month were 4266.53, 10426.14, 26185.33 and 30146.20 respectively. The average RFU measurements obtained for the ISUSN5 mutant plants were 33650.30, 18455.63, 31502.64 and 29204.34 respectively. E46K12 mutant plant also had the following RFU measurements; 13266.84, 26824, 13600.38 and 20204.88 respectively. Therefore, sampling method 2 did provided some few evidence to support the hypohesis that ageing is a contributing factor in α - synuclein aggregation unlike the sampling method 1.
Master's thesis in Biological chemistry