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Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis
Mitochondrial metabolism is entirely dependent on the biosynthesis of the [4Fe-4S] clusters, which are part of the subunits of the respiratory chain. The mitochondrial late ISC pathway mediates the formation of these clusters from simpler [2Fe-2S] molecules and transfers them to client proteins. Her...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10578589/ https://www.ncbi.nlm.nih.gov/pubmed/37792908 http://dx.doi.org/10.1371/journal.ppat.1010773 |
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author | Motyčková, Alžběta Voleman, Luboš Najdrová, Vladimíra Arbonová, Lenka Benda, Martin Dohnálek, Vít Janowicz, Natalia Malych, Ronald Šuťák, Róbert Ettema, Thijs J. G. Svärd, Staffan Stairs, Courtney W. Doležal, Pavel |
author_facet | Motyčková, Alžběta Voleman, Luboš Najdrová, Vladimíra Arbonová, Lenka Benda, Martin Dohnálek, Vít Janowicz, Natalia Malych, Ronald Šuťák, Róbert Ettema, Thijs J. G. Svärd, Staffan Stairs, Courtney W. Doležal, Pavel |
author_sort | Motyčková, Alžběta |
collection | PubMed |
description | Mitochondrial metabolism is entirely dependent on the biosynthesis of the [4Fe-4S] clusters, which are part of the subunits of the respiratory chain. The mitochondrial late ISC pathway mediates the formation of these clusters from simpler [2Fe-2S] molecules and transfers them to client proteins. Here, we characterized the late ISC pathway in one of the simplest mitochondria, mitosomes, of the anaerobic protist Giardia intestinalis that lost the respiratory chain and other hallmarks of mitochondria. In addition to IscA2, Nfu1 and Grx5 we identified a novel BolA1 homologue in G. intestinalis mitosomes. It specifically interacts with Grx5 and according to the high-affinity pulldown also with other core mitosomal components. Using CRISPR/Cas9 we were able to establish full bolA1 knock out, the first cell line lacking a mitosomal protein. Despite the ISC pathway being the only metabolic role of the mitosome no significant changes in the mitosome biology could be observed as neither the number of the mitosomes or their capability to form [2Fe-2S] clusters in vitro was affected. We failed to identify natural client proteins that would require the [2Fe-2S] or [4Fe-4S] cluster within the mitosomes, with the exception of [2Fe-2S] ferredoxin, which is itself part of the ISC pathway. The overall uptake of iron into the cellular proteins remained unchanged as also observed for the grx5 knock out cell line. The pull-downs of all late ISC components were used to build the interactome of the pathway showing specific position of IscA2 due to its interaction with the outer mitosomal membrane proteins. Finally, the comparative analysis across Metamonada species suggested that the adaptation of the late ISC pathway identified in G. intestinalis occurred early in the evolution of this supergroup of eukaryotes. |
format | Online Article Text |
id | pubmed-10578589 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-105785892023-10-17 Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis Motyčková, Alžběta Voleman, Luboš Najdrová, Vladimíra Arbonová, Lenka Benda, Martin Dohnálek, Vít Janowicz, Natalia Malych, Ronald Šuťák, Róbert Ettema, Thijs J. G. Svärd, Staffan Stairs, Courtney W. Doležal, Pavel PLoS Pathog Research Article Mitochondrial metabolism is entirely dependent on the biosynthesis of the [4Fe-4S] clusters, which are part of the subunits of the respiratory chain. The mitochondrial late ISC pathway mediates the formation of these clusters from simpler [2Fe-2S] molecules and transfers them to client proteins. Here, we characterized the late ISC pathway in one of the simplest mitochondria, mitosomes, of the anaerobic protist Giardia intestinalis that lost the respiratory chain and other hallmarks of mitochondria. In addition to IscA2, Nfu1 and Grx5 we identified a novel BolA1 homologue in G. intestinalis mitosomes. It specifically interacts with Grx5 and according to the high-affinity pulldown also with other core mitosomal components. Using CRISPR/Cas9 we were able to establish full bolA1 knock out, the first cell line lacking a mitosomal protein. Despite the ISC pathway being the only metabolic role of the mitosome no significant changes in the mitosome biology could be observed as neither the number of the mitosomes or their capability to form [2Fe-2S] clusters in vitro was affected. We failed to identify natural client proteins that would require the [2Fe-2S] or [4Fe-4S] cluster within the mitosomes, with the exception of [2Fe-2S] ferredoxin, which is itself part of the ISC pathway. The overall uptake of iron into the cellular proteins remained unchanged as also observed for the grx5 knock out cell line. The pull-downs of all late ISC components were used to build the interactome of the pathway showing specific position of IscA2 due to its interaction with the outer mitosomal membrane proteins. Finally, the comparative analysis across Metamonada species suggested that the adaptation of the late ISC pathway identified in G. intestinalis occurred early in the evolution of this supergroup of eukaryotes. Public Library of Science 2023-10-04 /pmc/articles/PMC10578589/ /pubmed/37792908 http://dx.doi.org/10.1371/journal.ppat.1010773 Text en © 2023 Motyčková et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Motyčková, Alžběta Voleman, Luboš Najdrová, Vladimíra Arbonová, Lenka Benda, Martin Dohnálek, Vít Janowicz, Natalia Malych, Ronald Šuťák, Róbert Ettema, Thijs J. G. Svärd, Staffan Stairs, Courtney W. Doležal, Pavel Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis |
title | Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis |
title_full | Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis |
title_fullStr | Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis |
title_full_unstemmed | Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis |
title_short | Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis |
title_sort | adaptation of the late isc pathway in the anaerobic mitochondrial organelles of giardia intestinalis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10578589/ https://www.ncbi.nlm.nih.gov/pubmed/37792908 http://dx.doi.org/10.1371/journal.ppat.1010773 |
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