Difference between revisions of "Lemieux 2019 bioRxiv"

Latest revision as of 11:08, 27 November 2021

Lemieux H, Subarsky P, Doblander C, Wurm M, Troppmair J, Gnaiger E (2019) Mitochondrial respiratory function as an early biomarker of apoptosis induced by growth factor removal. bioRxiv doi: https://doi.org/10.1101/151480 .

» Version 2 (2019-06-11) in preparation, bioRxiv Preprint Version 1 Open Access

Lemieux Helene, Subarsky P, Doblander C, Wurm M, Troppmair J, Gnaiger Erich (2019) bioRxiv

Abstract: Remodeling of mitochondrial metabolism is implicated in progression of cancer. Conversely, however, mitochondrial dysfunction and signaling play key roles in the induction of cell death. Apoptosis is induced following interleukin 3 (IL-3) depletion in mouse pro-myeloid 32D cells. Molecular signals of cell death are absent in 32D cells after short-term IL-3 deprivation (8 h). We addressed the question if changes in mitochondrial function can be detected by high-resolution respirometry as an early event in the induction of apoptosis. Respiration of living 32D cells was suppressed by 10 to 55% following 8 h removal of IL-3, but remained more stable in 32D cells expressing the v-RAF oncogene related to CRAF. In 32D cells deprived of IL3, succinate-supported respiration did not decline significantly, but respiratory capacities of the NADH-pathway and the combined NADH- and succinate-linked (NS) pathway were decreased compared to cells grown in the presence of IL-3. This was consistent with respiratory control exerted by impaired Complex IV activity, since there was not even the slightest excess Complex IV capacity above NS-pathway capacity. In contrast, electron flow reached only 60% when supported by succinate alone through Complexes II, III and IV, and was therefore relatively insensitive to Complex IV injuries up to a threshold of 40 % inhibition. After IL-3 depletion respiration increased by 15% following addition of cytochrome c, which provides a marker of mitochondrial outer membrane leakage, thus indicating mitochondrial fragility. Our results highlight a novel link between the key mitogenic and survival kinase CRAF and mitochondrial energy homeostasis.

• Keywords: Mitochondrial respiration, OXPHOS, cytochrome c oxidase, apoptosis, CRAF, interleukin 3 • Bioblast editor: Gnaiger E • O2k-Network Lab: CA_Edmonton_Lemieux H, AT_Innsbruck_Oroboros, AT_Innsbruck_Gnaiger E

Preprints for Gentle Science

» Preprints for Gentle Science

In the spirit of COST Action MitoEAGLE WG1

Contribution to K-Regio MitoFit

Cited by

Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5^th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002

Labels: MiParea: Respiration Pathology: Cancer Stress:Cell death Organism: Mouse Tissue;cell: Blood cells Preparation: Intact cells, Permeabilized cells Enzyme: Complex IV;cytochrome c oxidase, Marker enzyme Regulation: Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, Threshold;excess capacity, Uncoupler, Q-junction effect Coupling state: LEAK, ROUTINE, OXPHOS, ET Pathway: N, S, CIV, NS, ROX HRR: Oxygraph-2k

MitoEAGLEPublication, BEC 2020.2

@@ Line 1: / Line 1: @@
 {{Publication
-|title=Lemieux H, Subarsky P, Doblander C, Wurm M, Troppmair J, Gnaiger E (2017) Impairment of mitochondrial respiratory function as an early biomarker of apoptosis induced by growth factor removal. bioXriv doi: https://doi.org/10.1101/151480 .
+|title=Lemieux H, Subarsky P, Doblander C, Wurm M, Troppmair J, Gnaiger E (2019) Mitochondrial respiratory function as an early biomarker of apoptosis induced by growth factor removal. bioRxiv doi: https://doi.org/10.1101/151480 .
-|info=[http://biorxiv.org/content/early/2017/06/19/151480 '''bioRxiv Preprint''' Open Access]
+|info=[http://www.bioblast.at/index.php/File:Lemieux_et_al_v-Raf-2019-05-28.pdf '''Version 2 (2019-06-11) in preparation'''], [http://biorxiv.org/content/early/2017/06/19/151480 '''bioRxiv Preprint Version 1''' Open Access]
-|authors=Lemieux H, Subarsky P, Doblander C, Wurm M, Troppmair J, Gnaiger E
+|authors=Lemieux Helene, Subarsky P, Doblander C, Wurm M, Troppmair J, Gnaiger Erich
-|year=2017
+|year=2019
-|journal=bioXriv
+|journal=bioRxiv
-|abstract=Intracellular signaling pathways not only control cell proliferation and survival, but also regulate the provision of cellular energy and building blocks through mitochondrial and non-mitochondrial metabolism. Wild-type and oncogenic RAF kinases have been shown to prevent apoptosis following the removal of interleukin 3 (IL-3) from mouse pro-myeloid 32D cells by reducing mitochondrial reactive oxygen species production. To study primary effects of RAF on mitochondrial energy metabolism, we applied high-resolution respirometry after short-term IL-3 deprivation (8 h), before 32D cells show detectable signs of cell death. Respiration in intact 32D cells was suppressed as an early event following removal of IL-3, but remained more stable in 32D cells expressing the v-RAF oncogene. In permeabilized 32D cells deprived of IL-3, respiratory capacities of the NADH-pathway, the convergent NADH&succinate-pathway, and Complex IV activity were decreased compared to cells grown in the presence of IL-3, whereas succinate-supported respiration remained unchanged, consistent with control by Complex IV. The apparent Complex IV excess capacity was zero above NADH&succinate-pathway capacity reconstituting tricarboxylic acid cycle function. In comparison, electron flow reached only 60% when supported by succinate alone through Complexes II, III and IV, and was therefore relatively insensitive to Complex IV injuries up to a threshold of 40% inhibition. A slight increase in respiration following addition of cytochrome ''c'', a marker of mitochondrial outer membrane leakage, was present after IL-3 depletion, indicating mitochondrial fragility. Our results highlight a novel link between the key mitogenic and survival kinase CRAF and mitochondrial energy homeostasis.
+|abstract=Remodeling of mitochondrial metabolism is implicated in progression of cancer. Conversely, however, mitochondrial dysfunction and signaling play key roles in the induction of cell death. Apoptosis is induced following interleukin 3 (IL-3) depletion in mouse pro-myeloid 32D cells. Molecular signals of cell death are absent in 32D cells after short-term IL-3 deprivation (8 h). We addressed the question if changes in mitochondrial function can be detected by high-resolution respirometry as an early event in the induction of apoptosis. Respiration of living 32D cells was suppressed by 10 to 55% following 8 h removal of IL-3, but remained more stable in 32D cells expressing the v-RAF oncogene related to CRAF. In 32D cells deprived of IL3, succinate-supported respiration did not decline significantly, but respiratory capacities of the NADH-pathway and the combined NADH- and succinate-linked (NS) pathway were decreased compared to cells grown in the presence of IL-3. This was consistent with respiratory control exerted by impaired Complex IV activity, since there was not even the slightest excess Complex IV capacity above NS-pathway capacity. In contrast, electron flow reached only 60% when supported by succinate alone through Complexes II, III and IV, and was therefore relatively insensitive to Complex IV injuries up to a threshold of 40 % inhibition. After IL-3 depletion respiration increased by 15% following addition of cytochrome ''c'', which provides a marker of mitochondrial outer membrane leakage, thus indicating mitochondrial fragility. Our results highlight a novel link between the key mitogenic and survival kinase CRAF and mitochondrial energy homeostasis.
+<br><br>
 |keywords=Mitochondrial respiration, OXPHOS, cytochrome c oxidase, apoptosis, CRAF, interleukin 3
-|editor=[[Gnaiger E]],
+|editor=[[Gnaiger E]]
-|mipnetlab=CA_Edmonton_Lemieux H, AT_Innsbruck_OROBOROS, AT_Innsbruck_Gnaiger E
+|mipnetlab=CA_Edmonton_Lemieux H, AT_Innsbruck_Oroboros, AT_Innsbruck_Gnaiger E
 }}
+== Preprints for Gentle Science ==
+::::» [[Gentle_Science#Preprints_for_Gentle_Science |Preprints for Gentle Science]]
+[[Image:MITOEAGLE-logo.jpg|60px|link=http://www.mitoglobal.org/index.php/MitoEAGLE|COST Action MitoEAGLE]] In the spirit of COST Action [[WG1_MitoEAGLE_protocols,_terminology,_documentation#Documentation |MitoEAGLE WG1]]
+[[Image:MitoFit.jpg|60px|link=http://www.mitofit.org/index.php/K-Regio MitoFit |MitoFit]] Contribution to [[K-Regio MitoFit]]
+== Cited by ==
+{{Template:Cited by Gnaiger 2020 BEC MitoPathways}}
 {{Labeling
 |area=Respiration
@@ Line 18: / Line 29: @@
 |preparations=Intact cells, Permeabilized cells
 |enzymes=Complex IV;cytochrome c oxidase, Marker enzyme
-|topics=Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, Q-junction effect, Threshold;excess capacity, Uncoupler
+|topics=Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, Threshold;excess capacity, Uncoupler, Q-junction effect
-|couplingstates=LEAK, ROUTINE, OXPHOS, ETS
+|couplingstates=LEAK, ROUTINE, OXPHOS, ET
 |pathways=N, S, CIV, NS, ROX
 |instruments=Oxygraph-2k
+|additional=MitoEAGLEPublication, BEC 2020.2
 }}