Ubiquitination Occurs in the Mitochondrial Matrix by Eclipsed Targeted Components of the Ubiquitination Machinery

Yu Zhang, Ofri Karmon, Koyeli Das, Reuven Wiener, Norbert Lehming, Ophry Pines*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations


Ubiquitination is a critical type of post-translational modification in eukaryotic cells. It is involved in regulating nearly all cellular processes in the cytosol and nucleus. Mitochondria, known as the metabolism heart of the cell, are organelles that evolved from bacteria. Using the subcellular compartment-dependent α-complementation, we detect multiple components of ubiquitination machinery as being eclipsed distributed to yeast mitochondria. Ubiquitin conjugates and mono-ubiquitin can be detected in lysates of isolated mitochondria from cells expressing HA-Ub and treated with trypsin. By expressing MTS (mitochondrial targeting sequence) targeted HA-tagged ubiquitin, we demonstrate that certain ubiquitination events specifically occur in yeast mitochondria and are independent of proteasome activity. Importantly, we show that the E2 Rad6 affects the pattern of protein ubiquitination in mitochondria and provides an in vivo assay for its activity in the matrix of the organelle. This study shows that ubiquitination occurs in the mitochondrial matrix by eclipsed targeted components of the ubiquitin machinery, providing a new perspective on mitochondrial and ubiquitination research.

Original languageAmerican English
Article number4109
Issue number24
StatePublished - 17 Dec 2022

Bibliographical note

Publisher Copyright:
© 2022 by the authors.


  • E2
  • Rad6
  • dual targeting
  • eclipsed distribution
  • mitochondrial matrix
  • ubiquitination


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