Disrupting the plastidic iron-sulfur cluster biogenesis pathway in Toxoplasma gondii has pleiotropic effects irreversibly impacting parasite viability

Renaud, Eléa A; Pamukcu, Sarah; Cerutti, Aude; Berry, Laurence; Lemaire-Vieille, Catherine; Yamaryo‐Botté, Yoshiki; Botté, Cyrille Y.; Besteiro, Sébastien

doi:10.1016/j.jbc.2022.102243

Cited by 20 publications

(15 citation statements)

References 90 publications

(172 reference statements)

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“…Proteins were stained with primary antibodies for 1h, followed by three washes with PBS before incubation with secondary antibodies in a 1% BSA/PBS solution for 1h. Primary antibodies used in this study and their respective dilutions were rabbit polyclonal anti-IMC3 23 antibody diluted at 1/1,000, mouse monoclonal anti-SAG1 24 diluted at 1/1,000 (T3 1E5), mouse monoclonal anti-P21 25 diluted at 1/200 (T8 4G10), mouse monoclonal anti-F1-ATPase beta subunit diluted at 1/1,000 (gift of P. Bradley) and rabbit polyclonal anti-pyruvate dehydrogenase E2 subunit 26 diluted at 1/500. Cyst walls were stained with a 1/300 dilution of a biotin-labelled Dolichos biflorus lectin (L-6533, Sigma-Aldrich) for 1h and revealed using a 1/300 dilution of FITC-conjugated streptavidin (SNN1008, Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

“…We performed immunofluorescence assays (IFAs) on types I, II and III parasites treated for two days with 50 µM BPD to assess the effect of acute iron depletion on the integrity of the parasites. We used markers of the pellicle of the parasite (constituted by the plasma membrane and a double-membrane complex known as the inner membrane complex -IMC-) 29 and of the mitochondrion and apicoplast, two endosymbiotic organelles that host iron-containing proteins 8,9,26 . We observed important alterations of the pellicle, and problems of DNA or apicoplast segregation in daughter cells (Fig.…”

Section: Acute Iron Depletion Strongly Impacts the Parasite Ultrastru...mentioning

confidence: 99%

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Iron depletion has different consequences on the growth and survival ofToxoplasma gondiistrains

Renaud,

Maupin,

Bordat

et al. 2023

Preprint

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Toxoplasma gondiiis an obligate intracellular parasite that is responsible for a pathology called toxoplasmosis which is primarily affecting immunocompromised individuals and developing fetuses. The parasite is able to scavenge essential nutrients from its host to support its own growth and survival. Among them, iron is one of the most important elements needed to sustain basic cellular functions, as it is involved in a number of key metabolic processes, including oxygen transport, redox balance and electron transport. We have evaluated the effects of an iron chelator on the development of several parasite strains and found that they differed in their ability to tolerate iron depletion. The growth of parasites usually associated with a model of acute toxoplasmosis was strongly impacted by iron depletion, while cystogenic strains were less sensitive as they were able to convert into persisting developmental forms which are associated with the chronic form of the disease. Ultrastructural and biochemical characterization of the impact of iron depletion on the parasites also highlighted striking changes in both in their metabolism and the one of the host, with a marked accumulation of lipid droplets and perturbation of lipid homeostasis. Overall, our study demonstrates that although acute iron depletion has an important effect on the growth ofT. gondii, it has a more profound impact on actively dividing parasites, while less metabolically-active parasite forms may be able to avoid some of the most detrimental consequences.

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Section: Methodsmentioning

confidence: 99%

Section: Acute Iron Depletion Strongly Impacts the Parasite Ultrastru...mentioning

confidence: 99%

Iron depletion has different consequences on the growth and survival ofToxoplasma gondiistrains

Renaud,

Maupin,

Bordat

et al. 2023

Preprint

Self Cite

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“…The apicoplast is an essential organelle required for fatty acid, isoprenoid, iron-sulfur cluster and heme synthesis [2][3][4][5][6][7][8]. Genetic manipulations or perturbation of the apicoplast with pharmacological compounds results in apicoplast inheritance defects and subsequently in parasite death [9][10][11].…”

Section: Introductionmentioning

confidence: 99%

F-actin and Myosin F control apicoplast elongation dynamics which drive apicoplast-centrosome association inToxoplasma gondii

Devarakonda

Sarmiento

Heaslip

2023

Preprint

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Toxoplasma gondiicontains an essential single plastid organelle known as the apicoplast that is necessary for fatty acid, isoprenoid, and heme synthesis. Perturbations affecting apicoplast function leads to parasite death. To maintain a functional apicoplast, the parasite must execute two important cellular processes: accurate division of this single copy organelle into daughter parasites during cell division and trafficking of nuclear encoded apicoplast proteins (NEAT). In this study we demonstrate that F-actin and an unconventional myosin motor, TgMyoF, are important for both processes. Live cell imaging demonstrates that during division the apicoplast is highly dynamic, exhibiting branched, U-shaped and linear morphologies that are dependent on TgMyoF and actin. These dynamics appear to control apicoplast association with the centrosome and positioning of the centrosome at the apicoplast tips. Loss of apicoplast dynamics correlated with reduced apicoplast-centrosome association and ultimately apicoplast inheritance defects. In addition, we uncovered the role of TgMyoF and actin in NEAT protein trafficking. Vesicles containing the apicoplast protein APT1 were only observed during apicoplast division in control parasites, however loss of TgMyoF and actin lead to accumulation of vesicles in the cytosol, with only a small impact on vesicle movement suggesting that this actomyosin system is important for vesicle fusion with the apicoplast. Consequently, loss of TgMyoF resulted in reduced apicoplast length. This study has provided crucial new insight into mechanisms and timing of protein trafficking to the apicoplast and demonstrated how apicoplast-centrosome association, a key step in the apicoplast division cycle, is control by the actomyosin cytoskeleton.

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“…Notably, for the latter two processes, it remains to be experimentally demonstrated whether direct electron transfer reactions from FDX to their targets are involved or whether the FDX dependence occurs indirectly via a putative FDX function in apicoplast Fe/S protein biogenesis [19]. In both plastids and apicoplasts, this process is catalysed by the bacteria-inherited SUF system [21,22] for which an involvement of a FDX has not been documented yet [23], but in similarity to mitochondrial Fe/S protein biogenesis (see below) an electron donor seems mechanistically plausible. The plant-type FDX proteins will not be further discussed here.…”

Section: Introductionmentioning

confidence: 99%

Mitochondrial [2Fe‐2S] ferredoxins: new functions for old dogs

et al. 2022

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Ferredoxins (FDXs) comprise a large family of iron–sulfur proteins that shuttle electrons from NADPH and FDX reductases into diverse biological processes. This review focuses on the structure, function and specificity of mitochondrial [2Fe‐2S] FDXs that are related to bacterial FDXs due to their endosymbiotic inheritance. Their classical function in cytochrome P450‐dependent steroid transformations was identified around 1960, and is exemplified by mammalian FDX1 (aka adrenodoxin). Thirty years later the essential function in cellular Fe/S protein biogenesis was discovered for the yeast mitochondrial FDX Yah1 that is additionally crucial for the formation of haem a and ubiquinone CoQ6. In mammals, Fe/S protein biogenesis is exclusively performed by the FDX1 paralog FDX2, despite the high structural similarity of both proteins. Recently, additional and specific roles of human FDX1 in haem a and lipoyl cofactor biosyntheses were described. For lipoyl synthesis, FDX1 transfers electrons to the radical S‐adenosyl methionine‐dependent lipoyl synthase to kickstart its radical chain reaction. The high target specificity of the two mammalian FDXs is contained within small conserved sequence motifs, that upon swapping change the target selection of these electron donors.

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Disrupting the plastidic iron-sulfur cluster biogenesis pathway in Toxoplasma gondii has pleiotropic effects irreversibly impacting parasite viability

Cited by 20 publications

References 90 publications

Iron depletion has different consequences on the growth and survival ofToxoplasma gondiistrains

Iron depletion has different consequences on the growth and survival ofToxoplasma gondiistrains

F-actin and Myosin F control apicoplast elongation dynamics which drive apicoplast-centrosome association inToxoplasma gondii

Mitochondrial [2Fe‐2S] ferredoxins: new functions for old dogs

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