Mitochondrial inner membrane protein (mitofilin) knockdown induces cell death by apoptosis via an AIF-PARP-dependent mechanism and cell cycle arrest

American Journal of Physiology. Cell Physiology
Ngonidzashe B MadungweJean C Bopassa

Abstract

Mitofilin is an inner membrane protein that has been defined as a mitochondria-shaping protein in controlling and maintaining mitochondrial cristae structure and remodeling. We determined the role of mitofilin in cell survival by investigating the mechanism underlying mitofilin knockdown-induced cell death by apoptosis. Cultured H9c2 myoblasts and HEK 293 cells were treated with mitofilin siRNA or scrambled siRNA for 24 h. Cell death (apoptosis), caspase 3 activity and cell cycle phases were assessed by flow cytometry, while cytochrome c release and intracellular ATP production were measured by ELISA. Mitofilin, apoptosis-inducing factor (AIF) and poly(ADP-ribose) polymerase (PARP) expression were measured by Western blot analysis and calpain activity was assessed using a calpain activity kit. Mitochondrial images were taken using electron microscopy. We found that mitofilin knockdown increases apoptosis mainly via activation of the AIF-PARP pathway leading to nuclear fragmentation that is correlated with S phase arrest of the cell cycle. Knockdown of mitofilin also led to mitochondrial swelling and damage of cristae that is associated with the increase in reactive oxygen species production and mitochondrial calpain activity, a...Continue Reading

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Citations

Sep 28, 2018·Journal of Cellular Physiology·Yansheng FengJean C Bopassa
Oct 5, 2019·Thoracic Cancer·Yasuhiro HiyoshiKatsuhiko Naoki
Dec 12, 2019·Journal of Biochemistry·Matthew J EramoMichael T Ryan
Feb 12, 2019·Frontiers in Neuroscience·Victor S Van LaarSarah B Berman
Dec 11, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Roberta MacrìVincenzo Mollace
Sep 11, 2019·Clinical Neurology and Neurosurgery·Monserrat Pérez-RamírezNormand García-Hernández
Jul 25, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Valentina Lo GalboAntonella D'Anneo

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Methods Mentioned

BETA
transgenic
transfection
electrophoresis
ELISA
protein assay
Assay
flow cytometry
electron microscopy
flow-cytometry
nuclear translocation

Software Mentioned

SPSS
MICOS

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