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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., 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.

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Cruz-Zaragoza, L. D., Author
Dennerlein, S., Author
Linden, A.1, Author           
Yousefi, R., Author
Lavdovskaia, E., Author
Aich, A., Author
Falk, R. R., Author
Gomkale, R., Author
Schöndorf, T., Author
Bohnsack, M. T., Author
Richter-Dennerlein, R., Author
Urlaub, H.1, Author           
Rehling, P.2, Author           
Affiliations:
1Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society, ou_578613              
2Max Planck Fellow Peter Rehling, ou_1298545              

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 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.

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Language(s): eng - English
 Dates: 2021-10-202021-11-11
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1016/j.cell.2021.09.033
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Title: Cell
Source Genre: Journal
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Pages: - Volume / Issue: 184 (23) Sequence Number: - Start / End Page: 5824 - 5837.e15 Identifier: ISSN: 00928674