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@ARTICLE{Bogorodskiy:893931,
author = {Bogorodskiy, Andrey and Okhrimenko, Ivan and Maslov, Ivan
and Maliar, Nina and Burkatovskii, Dmitrii and von Ameln,
Florian and Schulga, Alexey and Jakobs, Philipp and
Altschmied, Joachim and Haendeler, Judith and Katranidis,
Alexandros and Sorokin, Ivan and Mishin, Alexey and
Gordeliy, Valentin and Büldt, Georg and Voos, Wolfgang and
Gensch, Thomas and Borshchevskiy, Valentin},
title = {{A}ccessing {M}itochondrial {P}rotein {I}mport in {L}iving
{C}ells by {P}rotein {M}icroinjection},
journal = {Frontiers in cell and developmental biology},
volume = {9},
issn = {2296-634X},
address = {Lausanne},
publisher = {Frontiers Media},
reportid = {FZJ-2021-02941},
pages = {698658},
year = {2021},
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.},
cin = {IBI-1 / IBI-6 / IBI-7},
ddc = {570},
cid = {I:(DE-Juel1)IBI-1-20200312 / I:(DE-Juel1)IBI-6-20200312 /
I:(DE-Juel1)IBI-7-20200312},
pnm = {5243 - Information Processing in Distributed Systems
(POF4-524) / 5352 - Understanding the Functionality of Soft
Matter and Biomolecular Systems (POF4-535) / 5241 -
Molecular Information Processing in Cellular Systems
(POF4-524)},
pid = {G:(DE-HGF)POF4-5243 / G:(DE-HGF)POF4-5352 /
G:(DE-HGF)POF4-5241},
typ = {PUB:(DE-HGF)16},
pubmed = {34307376},
UT = {WOS:000674898900001},
doi = {10.3389/fcell.2021.698658},
url = {https://juser.fz-juelich.de/record/893931},
}