ABSTRACT
Mitochondria and lysosomes are functionally linked, and their interdependent decline is a hallmark of aging and disease. Despite the long-standing connection between these organelles, the function(s) of lysosomes required to sustain mitochondrial health remains unclear. Here, working in yeast, we show that the lysosome-like vacuole maintains mitochondrial respiration by spatially compartmentalizing amino acids. Defects in vacuole function result in a breakdown in intracellular amino acid homeostasis, which drives age-related mitochondrial decline. Among amino acids, we find that cysteine is most toxic for mitochondria and show that elevated non-vacuolar cysteine impairs mitochondrial respiration by limiting intracellular iron availability through an oxidant-based mechanism. Cysteine depletion or iron supplementation restores mitochondrial health in vacuole-impaired cells and prevents mitochondrial decline during aging. These results demonstrate that cysteine toxicity is a major driver of age-related mitochondrial deterioration and identify vacuolar amino acid compartmentation as a cellular strategy to minimize amino acid toxicity.
Subject(s)
Cysteine/toxicity , Iron/metabolism , Mitochondria/metabolism , Amino Acids/metabolism , Cellular Senescence/physiology , Cysteine/metabolism , Homeostasis , Lysosomes/metabolism , Mitochondria/physiology , Mitophagy/physiology , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Vacuolar Proton-Translocating ATPases/metabolism , Vacuoles/metabolismABSTRACT
Mitochondrial decline is a hallmark of aging, and cells are equipped with many systems to regulate mitochondrial structure and function in response to stress and metabolic alterations. Here, using budding yeast, we identify a proteolytic pathway that contributes to alterations in mitochondrial structure in aged cells through control of the mitochondrial fusion GTPase Fzo1. We show that mitochondrial fragmentation in old cells correlates with reduced abundance of Fzo1, which is triggered by functional alterations in the vacuole, a known early event in aging. Fzo1 degradation is mediated by a proteolytic cascade consisting of the E3 ubiquitin ligases SCFMdm30 and Rsp5, and the Cdc48 cofactor Doa1. Fzo1 proteolysis is activated by metabolic stress that arises from vacuole impairment, and loss of Fzo1 degradation severely impairs mitochondrial structure and function. Together, these studies identify a new mechanism for stress-responsive regulation of mitochondrial structure that is activated during cellular aging.
Subject(s)
Endosomal Sorting Complexes Required for Transport/metabolism , F-Box Proteins/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Ubiquitin-Protein Ligase Complexes/metabolism , Adaptor Proteins, Signal Transducing/metabolism , Cellular Senescence/physiology , GTP Phosphohydrolases/metabolism , Membrane Fusion/physiology , Membrane Proteins/metabolism , Mitochondria/metabolism , Mitochondria/physiology , Mitochondrial Dynamics , Mitochondrial Proteins/metabolism , SKP Cullin F-Box Protein Ligases/metabolism , Saccharomyces cerevisiae/metabolism , Vacuoles/metabolismABSTRACT
A new device for isolating large quantities of old yeast cells expands the experimental boundaries of aging research.