An in vitro system to silence mitochondrial gene expression

2021 | journal article; research paper. A publication with affiliation to the University of Göttingen.

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​An in vitro system to silence mitochondrial gene expression​
Cruz-Zaragoza, L. D.; Dennerlein, S. ; Linden, A.; Yousefi, R.; Lavdovskaia, E.; Aich, A.   & Falk, R. R. et al.​ (2021) 
Cell184(23) pp. 5824​-5837.e15​.​ DOI: 

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Cruz-Zaragoza, Luis Daniel; Dennerlein, Sven ; Linden, Andreas; Yousefi, Roya; Lavdovskaia, Elena; Aich, Abhishek ; Falk, Rebecca R.; Gomkale, Ridhima; Schöndorf, Thomas; Bohnsack, Markus T. ; Rehling, Peter 
The human mitochondrial genome encodes thirteen core subunits of the oxidative phosphorylation system, and defects in mitochondrial gene expression lead to severe neuromuscular disorders. However, the mechanisms of mitochondrial gene expression remain poorly understood due to a lack of experimental approaches to analyze these processes. Here, we present an in vitro system to silence translation in purified mitochondria. In vitro import of chemically synthesized precursor-morpholino hybrids allows us to target translation of individual mitochondrial mRNAs. By applying this approach, we conclude that the bicistronic, overlapping ATP8/ATP6 transcript is translated through a single ribosome/mRNA engagement. We show that recruitment of COX1 assembly factors to translating ribosomes depends on nascent chain formation. By defining mRNA-specific interactomes for COX1 and COX2, we reveal an unexpected function of the cytosolic oncofetal IGF2BP1, an RNA-binding protein, in mitochondrial translation. Our data provide insight into mitochondrial translation and innovative strategies to investigate mitochondrial gene expression.
Issue Date
EXC 2067: Multiscale Bioimaging 
SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente 
Working Group
RG M. Bohnsack (Molecular Biology) 
RG Rehling (Mitochondrial Protein Biogenesis) 
RG Richter-Dennerlein (Mitoribosome Assembly) 
RG Urlaub (Bioanalytische Massenspektrometrie) 
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