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Journal Article

An in vitro system to silence mitochondrial gene expression

MPS-Authors
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Linden,  A.
Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society;

/persons/resource/persons15947

Urlaub,  H.
Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society;

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Citation

Cruz-Zaragoza, L. D., Dennerlein, S., Linden, A., Yousefi, R., Lavdovskaia, E., Aich, A., et al. (2021). An in vitro system to silence mitochondrial gene expression. Cell, 184(23), 5824-5837.e15. doi:10.1016/j.cell.2021.09.033.


Cite as: https://hdl.handle.net/21.11116/0000-000A-7D74-8
Abstract
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 mech-
anisms of mitochondrial gene expression remain poorly understood due to a lack of experimental ap-
proaches 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.