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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 and Ettema, Thijs J G , et al. (2023) In PLoS Pathogens 19(10).
Abstract

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... (More)

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 this of supergroup of eukaryotes.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
PLoS Pathogens
volume
19
issue
10
article number
e1010773
pages
24 pages
publisher
Public Library of Science (PLoS)
external identifiers
  • scopus:85173952251
  • pmid:37792908
ISSN
1553-7374
DOI
10.1371/journal.ppat.1010773
language
English
LU publication?
yes
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Copyright: © 2023 Motyčková et al. 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 credited.
id
1fda3074-a622-494d-8008-6dd44f707bec
date added to LUP
2023-10-16 11:11:36
date last changed
2024-04-19 02:22:24
@article{1fda3074-a622-494d-8008-6dd44f707bec,
  abstract     = {{<p>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 this of supergroup of eukaryotes.</p>}},
  author       = {{Motyčková, Alžběta and Voleman, Luboš and Najdrová, Vladimíra and Arbonová, Lenka and Benda, Martin and Dohnálek, Vít and Janowicz, Natalia and Malych, Ronald and Šuťák, Róbert and Ettema, Thijs J G and Svärd, Staffan and Stairs, Courtney W and Doležal, Pavel}},
  issn         = {{1553-7374}},
  language     = {{eng}},
  month        = {{10}},
  number       = {{10}},
  publisher    = {{Public Library of Science (PLoS)}},
  series       = {{PLoS Pathogens}},
  title        = {{Adaptation of the late ISC pathway in the anaerobic mitochondrial organelles of Giardia intestinalis}},
  url          = {{http://dx.doi.org/10.1371/journal.ppat.1010773}},
  doi          = {{10.1371/journal.ppat.1010773}},
  volume       = {{19}},
  year         = {{2023}},
}