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A system for inducible mitochondria-specific protein degradation in vivo
A system for inducible mitochondria-specific protein degradation in vivo
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Length:
20 minutes
Released:
Dec 20, 2022
Format:
Podcast episode
Description
Link to bioRxiv paper:
http://biorxiv.org/cgi/content/short/2022.12.20.521030v1?rss=1
Authors: Sanyal, S., Kouznetsova, A., Bjorkegren, C.
Abstract:
Targeted protein degradation systems developed for eukaryotes employ cytoplasmic machineries to perform proteolysis. This has prevented mitochondria-specific analysis of genome maintaining proteins that localize to both mitochondria and nucleus. Here, we present an inducible mitochondria-specific protein degradation system in Saccharomyces cerevisiae based on the Mesoplasma florum Lon (mf-Lon) protease and its corresponding ssrA tag (called PDT). We show that mitochondrially targeted mf-Lon protease efficiently and selectively degrades a PDT- tagged reporter protein localized to the mitochondrial matrix. The degradation can be induced by depleting adenine from the medium and tuned by altering the promoter strength of the MF-LON gene. Finally, we demonstrate that mf-Lon degrades endogenous, dually localized proteins inside mitochondria. In summary, our system is an efficient tool for analysis of intricate mitochondria-nuclear crosstalk essential for proper mitochondrial function.
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Podcast created by Paper Player, LLC
http://biorxiv.org/cgi/content/short/2022.12.20.521030v1?rss=1
Authors: Sanyal, S., Kouznetsova, A., Bjorkegren, C.
Abstract:
Targeted protein degradation systems developed for eukaryotes employ cytoplasmic machineries to perform proteolysis. This has prevented mitochondria-specific analysis of genome maintaining proteins that localize to both mitochondria and nucleus. Here, we present an inducible mitochondria-specific protein degradation system in Saccharomyces cerevisiae based on the Mesoplasma florum Lon (mf-Lon) protease and its corresponding ssrA tag (called PDT). We show that mitochondrially targeted mf-Lon protease efficiently and selectively degrades a PDT- tagged reporter protein localized to the mitochondrial matrix. The degradation can be induced by depleting adenine from the medium and tuned by altering the promoter strength of the MF-LON gene. Finally, we demonstrate that mf-Lon degrades endogenous, dually localized proteins inside mitochondria. In summary, our system is an efficient tool for analysis of intricate mitochondria-nuclear crosstalk essential for proper mitochondrial function.
Copy rights belong to original authors. Visit the link for more info
Podcast created by Paper Player, LLC
Released:
Dec 20, 2022
Format:
Podcast episode
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