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Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery

The cytosolic iron–sulfur (Fe-S) cluster assembly (CIA) pathway delivers Fe-S clusters to nuclear and cytosolic Fe-S proteins involved in essential cellular functions. Although the delivery process is regulated by the availability of iron and oxygen, it remains unclear how CIA components orchestrate...

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Autores principales: Fan, Xiaorui, Barshop, William D., Vashisht, Ajay A., Pandey, Vijaya, Leal, Stephanie, Rayatpisheh, Shima, Jami-Alahmadi, Yasaman, Sha, Jihui, Wohlschlegel, James A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9243173/
https://www.ncbi.nlm.nih.gov/pubmed/35654137
http://dx.doi.org/10.1016/j.jbc.2022.102094
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author Fan, Xiaorui
Barshop, William D.
Vashisht, Ajay A.
Pandey, Vijaya
Leal, Stephanie
Rayatpisheh, Shima
Jami-Alahmadi, Yasaman
Sha, Jihui
Wohlschlegel, James A.
author_facet Fan, Xiaorui
Barshop, William D.
Vashisht, Ajay A.
Pandey, Vijaya
Leal, Stephanie
Rayatpisheh, Shima
Jami-Alahmadi, Yasaman
Sha, Jihui
Wohlschlegel, James A.
author_sort Fan, Xiaorui
collection PubMed
description The cytosolic iron–sulfur (Fe-S) cluster assembly (CIA) pathway delivers Fe-S clusters to nuclear and cytosolic Fe-S proteins involved in essential cellular functions. Although the delivery process is regulated by the availability of iron and oxygen, it remains unclear how CIA components orchestrate the cluster transfer under varying cellular environments. Here, we utilized a targeted proteomics assay for monitoring CIA factors and substrates to characterize the CIA machinery. We find that nucleotide-binding protein 1 (NUBP1/NBP35), cytosolic iron–sulfur assembly component 3 (CIAO3/NARFL), and CIA substrates associate with nucleotide-binding protein 2 (NUBP2/CFD1), a component of the CIA scaffold complex. NUBP2 also weakly associates with the CIA targeting complex (MMS19, CIAO1, and CIAO2B) indicating the possible existence of a higher order complex. Interactions between CIAO3 and the CIA scaffold complex are strengthened upon iron supplementation or low oxygen tension, while iron chelation and reactive oxygen species weaken CIAO3 interactions with CIA components. We further demonstrate that CIAO3 mutants defective in Fe-S cluster binding fail to integrate into the higher order complexes. However, these mutants exhibit stronger associations with CIA substrates under conditions in which the association with the CIA targeting complex is reduced suggesting that CIAO3 and CIA substrates may associate in complexes independently of the CIA targeting complex. Together, our data suggest that CIA components potentially form a metabolon whose assembly is regulated by environmental cues and requires Fe-S cluster incorporation in CIAO3. These findings provide additional evidence that the CIA pathway adapts to changes in cellular environment through complex reorganization.
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spelling pubmed-92431732022-07-01 Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery Fan, Xiaorui Barshop, William D. Vashisht, Ajay A. Pandey, Vijaya Leal, Stephanie Rayatpisheh, Shima Jami-Alahmadi, Yasaman Sha, Jihui Wohlschlegel, James A. J Biol Chem Research Article The cytosolic iron–sulfur (Fe-S) cluster assembly (CIA) pathway delivers Fe-S clusters to nuclear and cytosolic Fe-S proteins involved in essential cellular functions. Although the delivery process is regulated by the availability of iron and oxygen, it remains unclear how CIA components orchestrate the cluster transfer under varying cellular environments. Here, we utilized a targeted proteomics assay for monitoring CIA factors and substrates to characterize the CIA machinery. We find that nucleotide-binding protein 1 (NUBP1/NBP35), cytosolic iron–sulfur assembly component 3 (CIAO3/NARFL), and CIA substrates associate with nucleotide-binding protein 2 (NUBP2/CFD1), a component of the CIA scaffold complex. NUBP2 also weakly associates with the CIA targeting complex (MMS19, CIAO1, and CIAO2B) indicating the possible existence of a higher order complex. Interactions between CIAO3 and the CIA scaffold complex are strengthened upon iron supplementation or low oxygen tension, while iron chelation and reactive oxygen species weaken CIAO3 interactions with CIA components. We further demonstrate that CIAO3 mutants defective in Fe-S cluster binding fail to integrate into the higher order complexes. However, these mutants exhibit stronger associations with CIA substrates under conditions in which the association with the CIA targeting complex is reduced suggesting that CIAO3 and CIA substrates may associate in complexes independently of the CIA targeting complex. Together, our data suggest that CIA components potentially form a metabolon whose assembly is regulated by environmental cues and requires Fe-S cluster incorporation in CIAO3. These findings provide additional evidence that the CIA pathway adapts to changes in cellular environment through complex reorganization. American Society for Biochemistry and Molecular Biology 2022-05-30 /pmc/articles/PMC9243173/ /pubmed/35654137 http://dx.doi.org/10.1016/j.jbc.2022.102094 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Article
Fan, Xiaorui
Barshop, William D.
Vashisht, Ajay A.
Pandey, Vijaya
Leal, Stephanie
Rayatpisheh, Shima
Jami-Alahmadi, Yasaman
Sha, Jihui
Wohlschlegel, James A.
Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery
title Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery
title_full Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery
title_fullStr Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery
title_full_unstemmed Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery
title_short Iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery
title_sort iron-regulated assembly of the cytosolic iron–sulfur cluster biogenesis machinery
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9243173/
https://www.ncbi.nlm.nih.gov/pubmed/35654137
http://dx.doi.org/10.1016/j.jbc.2022.102094
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