Journal Article

FZJ-2021-02941

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Accessing Mitochondrial Protein Import in Living Cells by Protein Microinjection

Bogorodskiy, A. ; Okhrimenko, I. ; Maslov, I. ; Maliar, N. ; Burkatovskii, D. ; von Ameln, F. ; Schulga, A. ; Jakobs, P. ; Altschmied, J. ; Haendeler, J. ; Katranidis, A.FZJ* ; Sorokin, I. ; Mishin, A. ; Gordeliy, V.FZJ* ; Büldt, G. ; Voos, W. ; Gensch, T.FZJ* ; Borshchevskiy, V. (Corresponding author)FZJ*

2021
Frontiers Media Lausanne

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Please use a persistent id in citations: http://hdl.handle.net/2128/28108  doi:10.3389/fcell.2021.698658

Abstract: Mitochondrial protein biogenesis relies almost exclusively on the expression of nuclear-encoded polypeptides. The current model postulates that most of these proteins have to be delivered to their final mitochondrial destination after their synthesis in the cytoplasm. However, the knowledge of this process remains limited due to the absence of proper experimental real-time approaches to study mitochondria in their native cellular environment. We developed a gentle microinjection procedure for fluorescent reporter proteins allowing a direct non-invasive study of protein transport in living cells. As a proof of principle, we visualized potential-dependent protein import into mitochondria inside intact cells in real-time. We validated that our approach does not distort mitochondrial morphology and preserves the endogenous expression system as well as mitochondrial protein translocation machinery. We observed that a release of nascent polypeptides chains from actively translating cellular ribosomes by puromycin strongly increased the import rate of the microinjected pre-protein. This suggests that a substantial amount of mitochondrial translocase complexes was involved in co-translational protein import of endogenously expressed pre-proteins. Our protein microinjection method opens new possibilities to study the role of mitochondrial protein import in cell models of various pathological conditions as well as aging processes.

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Contributing Institute(s):

1. Molekular- und Zellphysiologie (IBI-1)
2. Zelluläre Strukturbiologie (IBI-6)
3. Strukturbiochemie (IBI-7)

Research Program(s):

1. 5243 - Information Processing in Distributed Systems (POF4-524) (POF4-524)
2. 5352 - Understanding the Functionality of Soft Matter and Biomolecular Systems (POF4-535) (POF4-535)
3. 5241 - Molecular Information Processing in Cellular Systems (POF4-524) (POF4-524)

Appears in the scientific report 2021

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Database coverage:
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