Down-regulation of the mitochondrial matrix peptidase ClpP in muscle cells causes mitochondrial dysfunction and decreases cell proliferation
- PMID: 26721594
- PMCID: PMC5584630
- DOI: 10.1016/j.freeradbiomed.2015.12.021
Down-regulation of the mitochondrial matrix peptidase ClpP in muscle cells causes mitochondrial dysfunction and decreases cell proliferation
Abstract
The caseinolytic peptidase P (ClpP) is the endopeptidase component of the mitochondrial matrix ATP-dependent ClpXP protease. ClpP degrades unfolded proteins to maintain mitochondrial protein homeostasis and is involved in the initiation of the mitochondrial unfolded protein response (UPR(mt)). Outside of an integral role in the UPR(mt), the cellular function of ClpP is not well characterized in mammalian cells. To investigate the role of ClpP in mitochondrial function, we generated C2C12 muscle cells that are deficient in ClpP using siRNA or stable knockdown using lentiviral transduction. Reduction of ClpP levels by ~70% in C2C12 muscle cells resulted in a number of mitochondrial alterations including reduced mitochondrial respiration and reduced oxygen consumption rate in response to electron transport chain (ETC) complex I and II substrates. The reduction in ClpP altered mitochondrial morphology, changed the expression level of mitochondrial fission protein Drp1 and blunted UPR(mt) induction. In addition, ClpP deficient cells showed increased generation of reactive oxygen species (ROS) and decreased membrane potential. At the cellular level, reduction of ClpP impaired myoblast differentiation, cell proliferation and elevated phosphorylation of eukaryotic initiation factor 2 alpha (eIF2α) suggesting an inhibition of translation. Our study is the first to define the effects of ClpP deficiency on mitochondrial function in muscle cells in vitro. In addition, we have uncovered novel effects of ClpP on mitochondrial morphology, cell proliferation and protein translation pathways in muscle cells.
Keywords: ClpP; ClpX; Mitochondrial fission/fusion; Mitochondrial unfolded protein response; Reactive oxygen species; Respiration.
Copyright © 2015 Elsevier Inc. All rights reserved.
Figures
![Fig. 1](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/5584630/bin/nihms894452f1.gif)
![Fig. 2](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/5584630/bin/nihms894452f2.gif)
![Fig. 3](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/5584630/bin/nihms894452f3.gif)
![Fig. 4](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/5584630/bin/nihms894452f4.gif)
![Fig. 5](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/5584630/bin/nihms894452f5.gif)
![Fig. 6](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/5584630/bin/nihms894452f6.gif)
![Fig. 7](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/5584630/bin/nihms894452f7.gif)
Similar articles
-
Senescent Hepatocytes in Decompensated Liver Show Reduced UPRMT and Its Key Player, CLPP, Attenuates Senescence In Vitro.Cell Mol Gastroenterol Hepatol. 2019;8(1):73-94. doi: 10.1016/j.jcmgh.2019.03.001. Epub 2019 Mar 13. Cell Mol Gastroenterol Hepatol. 2019. PMID: 30878663 Free PMC article.
-
Loss of mitochondrial protease ClpP protects mice from diet-induced obesity and insulin resistance.EMBO Rep. 2018 Mar;19(3):e45009. doi: 10.15252/embr.201745009. Epub 2018 Feb 2. EMBO Rep. 2018. PMID: 29420235 Free PMC article.
-
Glutamine depletion disrupts mitochondrial integrity and impairs C2C12 myoblast proliferation, differentiation, and the heat-shock response.Nutr Res. 2020 Dec;84:42-52. doi: 10.1016/j.nutres.2020.09.006. Epub 2020 Sep 19. Nutr Res. 2020. PMID: 33189431
-
Knockout Mouse Studies Show That Mitochondrial CLPP Peptidase and CLPX Unfoldase Act in Matrix Condensates near IMM, as Fast Stress Response in Protein Assemblies for Transcript Processing, Translation, and Heme Production.Genes (Basel). 2024 May 27;15(6):694. doi: 10.3390/genes15060694. Genes (Basel). 2024. PMID: 38927630 Free PMC article. Review.
-
Mitochondrial Caseinolytic Protease P: A Possible Novel Prognostic Marker and Therapeutic Target in Cancer.Int J Mol Sci. 2021 Jun 9;22(12):6228. doi: 10.3390/ijms22126228. Int J Mol Sci. 2021. PMID: 34207660 Free PMC article. Review.
Cited by
-
Mitochondrial stress response and myogenic differentiation.Front Cell Dev Biol. 2024 Apr 12;12:1381417. doi: 10.3389/fcell.2024.1381417. eCollection 2024. Front Cell Dev Biol. 2024. PMID: 38681520 Free PMC article. Review.
-
Intraneuronal β-amyloid impaired mitochondrial proteostasis through the impact on LONP1.Proc Natl Acad Sci U S A. 2023 Dec 19;120(51):e2316823120. doi: 10.1073/pnas.2316823120. Epub 2023 Dec 13. Proc Natl Acad Sci U S A. 2023. PMID: 38091289 Free PMC article.
-
From mitochondria to sarcopenia: role of 17β-estradiol and testosterone.Front Endocrinol (Lausanne). 2023 Apr 20;14:1156583. doi: 10.3389/fendo.2023.1156583. eCollection 2023. Front Endocrinol (Lausanne). 2023. PMID: 37152937 Free PMC article. Review.
-
Ssu72 phosphatase is essential for thermogenic adaptation by regulating cytosolic translation.Nat Commun. 2023 Feb 25;14(1):1097. doi: 10.1038/s41467-023-36836-y. Nat Commun. 2023. PMID: 36841836 Free PMC article.
-
Knowledge mapping of research on the mitochondrial unfolded protein response: a bibliometric and visual analysis.Ann Transl Med. 2023 Jan 31;11(2):64. doi: 10.21037/atm-22-6423. Epub 2023 Jan 13. Ann Transl Med. 2023. PMID: 36819568 Free PMC article.
References
-
- Lin MT, Beal MF. Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature. 2006;443:787–795. - PubMed
-
- Luce K, Weil AC, Osiewacz HD. Mitochondrial protein quality control systems in aging and disease. Adv Exp Med Biol. 2010;694:108–125. - PubMed
-
- Bota DA, Davies KJ. Lon protease preferentially degrades oxidized mitochondrial aconitase by an ATP-stimulated mechanism. Nat Cell Biol. 2002;4:674–680. - PubMed
-
- Truscott KN, Bezawork-Geleta A, Dougan DA. Unfolded protein responses in bacteria and mitochondria: A central role for the ClpXP machine. IUBMB Life. 2011;63:955–963. - PubMed
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous