MitoRibo-Tag Mice Provide a Tool for In Vivo Studies of Mitoribosome Composition

Cell Rep. 2019 Nov 5;29(6):1728-1738.e9. doi: 10.1016/j.celrep.2019.09.080.

Abstract

Mitochondria harbor specialized ribosomes (mitoribosomes) necessary for the synthesis of key membrane proteins of the oxidative phosphorylation (OXPHOS) machinery located in the mitochondrial inner membrane. To date, no animal model exists to study mitoribosome composition and mitochondrial translation coordination in mammals in vivo. Here, we create MitoRibo-Tag mice as a tool enabling affinity purification and proteomics analyses of mitoribosomes and their interactome in different tissues. We also define the composition of an assembly intermediate formed in the absence of MTERF4, necessary for a late step in mitoribosomal biogenesis. We identify the orphan protein PUSL1, which interacts with a large subunit assembly intermediate, and demonstrate that it is an inner-membrane-associated mitochondrial matrix protein required for efficient mitochondrial translation. This work establishes MitoRibo-Tag mice as a powerful tool to study mitoribosomes in vivo, enabling future studies on the mitoribosome interactome under different physiological states, as well as in disease and aging.

Keywords: MitoRibo-Tag mice; OXPHOS; in vivo mouse model; mitochondria; mitochondrial DNA; mitochondrial biogenesis; mitochondrial gene expression; mitochondrial ribosome; ribosome; translation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • GTP-Binding Proteins / genetics
  • GTP-Binding Proteins / metabolism
  • Heart / physiology
  • Kidney / metabolism
  • Liver / metabolism
  • Mice
  • Mice, Transgenic
  • Mitochondria / genetics
  • Mitochondria / metabolism*
  • Mitochondrial Membranes / metabolism*
  • Mitochondrial Proteins / genetics
  • Mitochondrial Proteins / metabolism*
  • Mitochondrial Ribosomes / metabolism*
  • Myocardium / metabolism
  • Protein Biosynthesis*
  • Protein Interaction Maps
  • Proteome / metabolism
  • Proteomics
  • Ribosomal Proteins / genetics
  • Ribosomal Proteins / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*

Substances

  • MTERF4 protein, mouse
  • Mitochondrial Proteins
  • Proteome
  • Ribosomal Proteins
  • Transcription Factors
  • GTP-Binding Proteins
  • GTPBP2 protein, mouse