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The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei

The mitochondrial F(0)F(1) ATP synthase is an essential multi-subunit protein complex in the vast majority of eukaryotes but little is known about its composition and role in Trypanosoma brucei, an early diverged eukaryotic pathogen. We purified the F(0)F(1) ATP synthase by a combination of affinity...

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Autores principales: Zíková, Alena, Schnaufer, Achim, Dalley, Rachel A., Panigrahi, Aswini K., Stuart, Kenneth D.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2674945/
https://www.ncbi.nlm.nih.gov/pubmed/19436713
http://dx.doi.org/10.1371/journal.ppat.1000436
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author Zíková, Alena
Schnaufer, Achim
Dalley, Rachel A.
Panigrahi, Aswini K.
Stuart, Kenneth D.
author_facet Zíková, Alena
Schnaufer, Achim
Dalley, Rachel A.
Panigrahi, Aswini K.
Stuart, Kenneth D.
author_sort Zíková, Alena
collection PubMed
description The mitochondrial F(0)F(1) ATP synthase is an essential multi-subunit protein complex in the vast majority of eukaryotes but little is known about its composition and role in Trypanosoma brucei, an early diverged eukaryotic pathogen. We purified the F(0)F(1) ATP synthase by a combination of affinity purification, immunoprecipitation and blue-native gel electrophoresis and characterized its composition and function. We identified 22 proteins of which five are related to F(1) subunits, three to F(0) subunits, and 14 which have no obvious homology to proteins outside the kinetoplastids. RNAi silencing of expression of the F(1) α subunit or either of the two novel proteins showed that they are each essential for the viability of procyclic (insect stage) cells and are important for the structural integrity of the F(0)F(1)-ATP synthase complex. We also observed a dramatic decrease in ATP production by oxidative phosphorylation after silencing expression of each of these proteins while substrate phosphorylation was not severely affected. Our procyclic T. brucei cells were sensitive to the ATP synthase inhibitor oligomycin even in the presence of glucose contrary to earlier reports. Hence, the two novel proteins appear essential for the structural organization of the functional complex and regulation of mitochondrial energy generation in these organisms is more complicated than previously thought.
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spelling pubmed-26749452009-05-15 The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei Zíková, Alena Schnaufer, Achim Dalley, Rachel A. Panigrahi, Aswini K. Stuart, Kenneth D. PLoS Pathog Research Article The mitochondrial F(0)F(1) ATP synthase is an essential multi-subunit protein complex in the vast majority of eukaryotes but little is known about its composition and role in Trypanosoma brucei, an early diverged eukaryotic pathogen. We purified the F(0)F(1) ATP synthase by a combination of affinity purification, immunoprecipitation and blue-native gel electrophoresis and characterized its composition and function. We identified 22 proteins of which five are related to F(1) subunits, three to F(0) subunits, and 14 which have no obvious homology to proteins outside the kinetoplastids. RNAi silencing of expression of the F(1) α subunit or either of the two novel proteins showed that they are each essential for the viability of procyclic (insect stage) cells and are important for the structural integrity of the F(0)F(1)-ATP synthase complex. We also observed a dramatic decrease in ATP production by oxidative phosphorylation after silencing expression of each of these proteins while substrate phosphorylation was not severely affected. Our procyclic T. brucei cells were sensitive to the ATP synthase inhibitor oligomycin even in the presence of glucose contrary to earlier reports. Hence, the two novel proteins appear essential for the structural organization of the functional complex and regulation of mitochondrial energy generation in these organisms is more complicated than previously thought. Public Library of Science 2009-05-15 /pmc/articles/PMC2674945/ /pubmed/19436713 http://dx.doi.org/10.1371/journal.ppat.1000436 Text en Zíková et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Zíková, Alena
Schnaufer, Achim
Dalley, Rachel A.
Panigrahi, Aswini K.
Stuart, Kenneth D.
The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei
title The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei
title_full The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei
title_fullStr The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei
title_full_unstemmed The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei
title_short The F(0)F(1)-ATP Synthase Complex Contains Novel Subunits and Is Essential for Procyclic Trypanosoma brucei
title_sort f(0)f(1)-atp synthase complex contains novel subunits and is essential for procyclic trypanosoma brucei
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2674945/
https://www.ncbi.nlm.nih.gov/pubmed/19436713
http://dx.doi.org/10.1371/journal.ppat.1000436
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