Coupling between the proteasomal catalytic unit and the 19S regulatory unit affects mitochondrial functionality after site-specific mutations of the 20S particle
DOI:
https://doi.org/10.11606/issn.1679-9836.v102iespe-204164Keywords:
Redox biology, Mitochondria, Proteasome, ProteolysisAbstract
A post-translational redox modification called S-glutathionylation in S. cerevisiae was described at Cys residues of the α5 subunit of the 20S catalytic unit of the proteasome, specifically α5-C76, posteriorly mutated to α5-C76S. The α5-C76S strain presented, as a phenotypic alteration, a higher frequency of the closed conformation of the catalytic chamber of the 20S unit and a shorter chronological life span (CLS: chronological life span). A double random mutation (DM: double mutated) in the α5 subunit (α5-S35P/C221S) induced the opening of the catalytic chamber and also increased CLS. This project aimed to assess the coupling between the catalytic and the regulatory units of the proteasome in some yeast strains (C76S, WT and DM) and to further evaluate their mitochondrial functionality. The study of the coupling of 20S-19S units was carried out in native gel electrophoresis. To determine mitochondrial functionality, the activity of citrate synthase was measured by the reaction between DTNB with CoA-SH. It was observed that in the C76S strain there was a lower degree of coupling between the 20S catalytic and 19S regulatory units, in addition to a decreased citrate synthase activity. Therefore, less coupling between the proteasomal units triggers mitochondrial dysfunction most likely due to deficient mitochondrial protein import, ultimately leading to decreased CLS, which can better our understanding in regards to the process of aging and premature cell death seen in degenerative disorders caused by protein accumulation.
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