Abstract
Mitophagy, the process that degrades mitochondria selectively through autophagy, is involved in the quality control of mitochondria in cells grown under respiratory conditions. In yeast, the presence of the Atg32 protein on the outer mitochondrial membrane allows for the recognition and targeting of superfluous or damaged mitochondria for degradation. Post-translational modifications such as phosphorylation are crucial for the execution of mitophagy. In our study we monitor the stability of Atg32 protein in the yeast S. cerevisiae and show that Atg32 is degraded under normal growth conditions, upon starvation or rapamycin treatment. The Atg32 turnover can be prevented by inhibition of the proteasome activity, suggesting that Atg32 is also ubiquitinated. Mass spectrometry analysis of purified Atg32 protein revealed that at least lysine residue in position 282 is ubiquitinated. Interestingly, the replacement of lysine 282 with alanine impaired Atg32 degradation only partially in the course of cell growth, suggesting that additional lysine residues on Atg32 might also be ubiquitinated. Our results provide the foundation to further elucidate the physiological significance of Atg32 turnover and the interplay between mitophagy and the proteasome.
Highlights
Mitochondria are organelles in charge of many crucial functions in cells
Mitophagy in yeast is induced during nitrogen starvation, during the stationary phase of growth, or through rapamycin treatment, where cells are grown in a strictly respiratory carbon source medium
These observations align with the data of Levchenko et al (2016), who showed that the C-terminal tagging of Atg32 by a ZZ tag did not interfere with mitophagy induced by nitrogen starvation or rapamycin treatment [12]
Summary
Mitochondria are organelles in charge of many crucial functions in cells. In eukaryotes, they are known to be the powerhouse of cells by producing ATP via oxidative phosphorylation; they are involved in different synthesis pathways such as the synthesis of certain amino acids and lipids and in signaling pathways, such as apoptosis and calcium. Mitochondrial dysfunctions have been associated with aging and with an increasing number of pathologies, including neurodegenerative diseases, cancer, and metabolic perturbation. Mitophagy is one of the processes involved in mitochondrial quality control. Mitophagy is a selective form of autophagy [1, 2]. Macroautophagy (hereafter called autophagy) is conserved among eukaryotic species and involves
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