Consequences of Cytochrome c Oxidase Assembly Defects for the Yeast Stationary Phase

Abstract

Functional assembly of cytochrome c oxidase (COX) is essential for an intact mitochondrial respiratory chain, although the consequences of a loss of assembled COX at yeast stationary phase remain largely unexplored. Stationary phase cultures are a good model for terminally differentiated cells in humans, such as neurons. Our goal is to define the physiological changes in mitochondrial respiration and oxidative stress between exponential and stationary phases in a respiratory competent strain and a set of COX assembly mutants with defects in a variety of aspects related to enzyme assembly. These results further our understanding of mitochondrial changes in terminally differentiated cells and aging cells. In this study, we show that a wild‐type respiratory competent yeast strain at stationary phase is characterized by a decreased oxidative capacity, as seen by a reduction in the amount of assembled COX, which is supported by a decrease in protein levels of several COX assembly factors. Wild‐type cells also have a disrupted mitochondrial reticulum during stationary phase. Loss of assembled COX results in the decreased abundance of many mitochondrial proteins at stationary phase, which we show is likely due to decreased membrane potential and changes in mitophagy. In addition to an altered mitochondrial proteome, COX assembly mutants display unexpected changes in markers of cellular oxidative stress at stationary phase. Specifically, differences in distribution of Sod1 between exponential and stationary phases give insight on location of reactive oxygen species (ROS) in the cell. Our results suggest that mitochondria may not be a major source of ROS at stationary phase in cells lacking an intact respiratory chain. Overall, we have identified fundamental differences between exponential and stationary phase mitochondria in respiratory competent yeast and have defined two distinct phenotypes, decreased levels of mitochondrial proteins and less mitochondrial oxidative damage, in cells with a defective respiratory chain. These findings suggest that COX deficiencies are associated with broader cellular phenotypes in terminally differentiated cells in humans.Support or Funding InformationNatural Sciences and Engineering Research Council of Canada (NSERC)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.