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

Research output: Contribution to journalArticlepeer-review

Abstract

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 languageEnglish
Article number4109
JournalCells
Volume11
Issue number24
DOIs
StatePublished - 17 Dec 2022

Keywords

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

All Science Journal Classification (ASJC) codes

  • General Biochemistry,Genetics and Molecular Biology

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