ISCA1 is essential for mitochondrial Fe4S4 biogenesis in vivo

Beilschmidt, Lena; Choudens, Sandrine Ollagnier de; Fournier, Marjorie; Sanakis, Ioannis; Hograindleur, Marc-André; Clémancey, Martin; Blondin, Geneviève; Schmucker, Stéphane; Eisenmann, Aurélie; Weiss, Amélie; Koebel, Pascale; Messaddeq, Nadia; Puccio, Hélène; Martelli, Alain

doi:10.1038/ncomms15124

Cited by 86 publications

(127 citation statements)

References 59 publications

(84 reference statements)

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“…The first studies on the mitochondrial [4Fe‐4S] cluster assembly step showed that three human proteins ISCA1, ISCA2, and IBA57 are required for this process . This functional association has been revisited in a recent work , which showed that ISCA2 and IBA57 are not required under standard physiological conditions for the maturation of mitochondrial [4Fe‐4S] proteins. Therefore, the so far available in vivo data do not provide a clear picture on how the three proteins operate in the cell for assembling a [4Fe‐4S] cluster.…”

Section: Discussionmentioning

confidence: 99%

A pathway for assembling [4Fe‐4S]²⁺ clusters in mitochondrial iron–sulfur protein biogenesis

et al. 2019

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During its late steps, the mitochondrial iron-sulfur cluster (ISC) assembly machinery leads to the formation of [4Fe-4S] clusters. In vivo studies revealed that several proteins are implicated in the biosynthesis and trafficking of [4Fe-4S] clusters in mitochondria. However, they do not provide a clear picture into how these proteins cooperate. Here, we showed that three late-acting components of the mitochondrial ISC assembly machinery (GLRX5, BOLA3, and NFU1) are part of a ISC assembly pathway leading to the synthesis of a [4Fe-4S] 2+ cluster on NFU1. We showed that the [2Fe-2S] 2+ GLRX5-BOLA3 complex transfers its cluster to monomeric apo NFU1 to form, in the presence of a reductant, a [4Fe-4S] 2+ cluster bound to dimeric NFU1. The cluster formation on NFU1 does not occur with [2Fe-2S] 2+ GLRX5, and thus, the [4Fe-4S] cluster assembly pathway is activated only in the presence of BOLA3. These results define NFU1 as an 'assembler' of [4Fe-4S] clusters, that is, a protein able of converting two [2Fe-2S] 2+ clusters into a [4Fe-4S] 2+ cluster. Finally, we found that the [4Fe-4S] 2+ cluster bound to NFU1 has a coordination site which is easily accessible to sulfur-containing ligands, as is typically observed in metallochaperones. This finding supports a role for NFU1 in promoting rapid and controlled cluster-exchange reaction.Abbreviations BMRB, biological magnetic resonance bank; Fe-S, iron-sulfur; GSH, reduced glutathione; HADDOCK, high ambiguity-driven protein-protein docking; HSQC, heteronuclear single quantum coherence; IPTG, isopropyl b-D-1-thiogalactopyranoside; ISC, iron-sulfur cluster; LB, Luria-Bertani; NOE, nuclear overhauser effect; OD, optical density; R 1 , longitudinal relaxation rate; R 2 , transverse relaxation rate; SAXS, smallangle X-ray scattering; SEC-MALS, size exclusion chromatography combined with multiangle light scattering; TCEP, tris (2-carboxyethyl) phosphine.

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Section: Discussionmentioning

confidence: 99%

A pathway for assembling [4Fe‐4S]²⁺ clusters in mitochondrial iron–sulfur protein biogenesis

et al. 2019

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“…Alternatively, overexpression of ISC2 in an ISCU1 knock-down background in mouse cells showed that ISCA1, but not ISCA2, was essential for the [4Fe–4S] cluster assembly, since ISCU2 could not rescue the [4Fe–4S] defect caused by silencing of ISCU1 [89]. Regardless, the two scaffold proteins in mouse and human interact with each other in vitro [89, 109], although only ISCU2 was able to bind IBA57 [89]. …”

Section: Fe–s Cluster Machineries: a Brief Overviewmentioning

confidence: 99%

Fe–S cluster assembly in the supergroup Excavata

Peña-Diaz

Lukeš

2018

J Biol Inorg Chem

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The majority of established model organisms belong to the supergroup Opisthokonta, which includes yeasts and animals. While enlightening, this focus has neglected protists, organisms that represent the bulk of eukaryotic diversity and are often regarded as primitive eukaryotes. One of these is the “supergroup” Excavata, which comprises unicellular flagellates of diverse lifestyles and contains species of medical importance, such as Trichomonas, Giardia, Naegleria, Trypanosoma and Leishmania. Excavata exhibits a continuum in mitochondrial forms, ranging from classical aerobic, cristae-bearing mitochondria to mitochondria-related organelles, such as hydrogenosomes and mitosomes, to the extreme case of a complete absence of the organelle. All forms of mitochondria house a machinery for the assembly of Fe–S clusters, ancient cofactors required in various biochemical activities needed to sustain every extant cell. In this review, we survey what is known about the Fe–S cluster assembly in the supergroup Excavata. We aim to bring attention to the diversity found in this group, reflected in gene losses and gains that have shaped the Fe–S cluster biogenesis pathways.

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“…ISCA1 knockout resulted in developmental block in embryos at 8.5 days and caused embryonic lethality (Figure ). The ISCA1 protein accepts Fe/S from a scaffold protein and transfers it to the respiratory complex . The Fe/S is necessary for the stability of the mitochondrial Fe/S containing proteins of the respiratory complex in eukaryotic cells .…”

Section: Discussionmentioning

confidence: 99%

“…The Fe/S is necessary for the stability of the mitochondrial Fe/S containing proteins of the respiratory complex in eukaryotic cells . ISCA1 knockdown by rAAV‐mediated shRNA resulted in a substantial decrease in Succinate Dehydrogenase Complex Iron Sulfur Subunit B (SDHB), NADH Dehydrogenase Fe‐S Protein 3 (NDUFS3) and NADH Dehydrogenase (Ubiquinone) Fe‐S Protein 5 (NDUFS5) in skeletal muscle . NDUFA9 is a Fe/S‐containing protein of the mitochondrial respiratory chain complex I and is essential for assembling and stabilizing complex I .…”

Section: Discussionmentioning

confidence: 99%

“…ISCA1 and ISCA2 behave as Fe/S carriers, accepting Fe/S from a scaffold protein and transferring it to target proteins, and function as important Fe/S cluster assembly machinery in mammalian cells. In HeLa cells, knockdown of ISCA1 and ISCA2 resulted in a decrease of the activities of mitochondrial Fe/S proteins, including aconitase, respiratory complex I, and lipoic acid synthase, through the ISC assembly pathway . ISCA1 and ISCA2 are the causal genes for multiple mitochondrial dysfunctions syndrome (MMDS).…”

Section: Introductionmentioning

confidence: 99%

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Knockout of ISCA1 causes early embryonic death in rats

Yang

Zhang

et al. 2019

Anim Models and Exp Med

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Background Iron‐sulfur cluster assembly 1 ( ISCA 1) is an iron‐sulfur (Fe/S) carrier protein that accepts Fe/S from a scaffold protein and transfers it to target proteins including the mitochondrial Fe/S containing proteins. ISCA 1 is also the newly identified causal gene for multiple mitochondrial dysfunctions syndrome ( MMDS ). However, our knowledge about the physiological function of ISCA 1 in vivo is currently limited. In this study, we generated an ISCA 1 knockout rat line and analyzed the embryo development. Methods ISCA 1 knockout rats were generated by replacing the exon1 of ISCA 1 gene with the mC herry‐Cre fusion gene using CRISPR ‐Cas9 technology. The ISCA 1 expression pattern was analyzed by fluorescence imaging using ISCA 1 promotor driven Cre and mC herry expression. The embryonic morphology was examinated by microscope and mitochondrial proteins were tested by Western blot. Results An ISCA 1 knockout rat line was obtained, which expressed mC herry‐Cre fusion protein. Both of the fluorescence images from mC herry and Cre induced mC herry in a reporter rat strain, showing that ISCA 1 expressed in most of the tissues in rats. The ISCA 1 knockout resulted in abnormal development at 8.5 days, with a significant decrease of NDUFA 9 protein and an increase of aconitase 2 ( ACO 2) in rat embryos. Conclusion Deletion of ISCA 1 induced early death in rats. ISCA 1 affected the expression of key proteins in the mitochondrial respiratory chain complex, suggesting that ISCA 1 has an important influence on the respiratory complex and energy metabolism.

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ISCA1 is essential for mitochondrial Fe4S4 biogenesis in vivo

Cited by 86 publications

References 59 publications

A pathway for assembling [4Fe‐4S]²⁺ clusters in mitochondrial iron–sulfur protein biogenesis

A pathway for assembling [4Fe‐4S]²⁺ clusters in mitochondrial iron–sulfur protein biogenesis

Fe–S cluster assembly in the supergroup Excavata

Knockout of ISCA1 causes early embryonic death in rats

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ISCA1 is essential for mitochondrial Fe4S4 biogenesis in vivo

Cited by 86 publications

References 59 publications

A pathway for assembling [4Fe‐4S]2+ clusters in mitochondrial iron–sulfur protein biogenesis

A pathway for assembling [4Fe‐4S]2+ clusters in mitochondrial iron–sulfur protein biogenesis

Fe–S cluster assembly in the supergroup Excavata

Knockout of ISCA1 causes early embryonic death in rats

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A pathway for assembling [4Fe‐4S]²⁺ clusters in mitochondrial iron–sulfur protein biogenesis

A pathway for assembling [4Fe‐4S]²⁺ clusters in mitochondrial iron–sulfur protein biogenesis