TY  - JOUR
AU  - Bogorodskiy, Andrey
AU  - Okhrimenko, Ivan
AU  - Maslov, Ivan
AU  - Maliar, Nina
AU  - Burkatovskii, Dmitrii
AU  - von Ameln, Florian
AU  - Schulga, Alexey
AU  - Jakobs, Philipp
AU  - Altschmied, Joachim
AU  - Haendeler, Judith
AU  - Katranidis, Alexandros
AU  - Sorokin, Ivan
AU  - Mishin, Alexey
AU  - Gordeliy, Valentin
AU  - Büldt, Georg
AU  - Voos, Wolfgang
AU  - Gensch, Thomas
AU  - Borshchevskiy, Valentin
TI  - Accessing Mitochondrial Protein Import in Living Cells by Protein Microinjection
JO  - Frontiers in cell and developmental biology
VL  - 9
SN  - 2296-634X
CY  - Lausanne
PB  - Frontiers Media
M1  - FZJ-2021-02941
SP  - 698658
PY  - 2021
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - 34307376
UR  - <Go to ISI:>//WOS:000674898900001
DO  - DOI:10.3389/fcell.2021.698658
UR  - https://juser.fz-juelich.de/record/893931
ER  -