GO:0140132 iron-sulfur cluster chaperone activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0140132 iron-sulfur cluster chaperone activity is a molecular function defined as binding to an iron-sulfur cluster and delivering it to an acceptor molecule [QuickGO].
• Iron-sulfur cluster chaperones are essential for the maturation of Fe-S proteins, which participate in electron transport, enzyme catalysis, and gene regulation [2, 4].
• Key chaperones include mitochondrial IscA1, cytosolic CIAO1, and the Hsp70 chaperone Ssq1p, which facilitate cluster transfer to scaffold and target proteins [2, 5, 8].
• Defects in Fe-S cluster chaperones are linked to human diseases such as neuromuscular disorders, cardiomyopathies, and cancer [1, 5].
• Studying GO:0140132 requires a combination of biochemical assays, CRISPR knockout models, and proteomic approaches to track cluster delivery [3, 7].
• EDITGENE provides CRISPR services to generate knockout, point-mutation, knock-in, and overexpression cell models for genes involved in iron-sulfur cluster chaperone activity.
Description
Iron-sulfur (Fe-S) clusters are ancient and versatile cofactors that are essential for the function of numerous proteins involved in electron transfer, substrate binding, and structural stabilization. The proper assembly and delivery of these clusters to target proteins rely on a dedicated machinery, including chaperones that bind Fe-S clusters and transfer them to acceptor molecules. This function is formally described by the Gene Ontology term GO:0140132, iron-sulfur cluster chaperone activity [QuickGO]. Understanding this activity is critical because Fe-S proteins are ubiquitous and their dysfunction is associated with a wide range of human diseases, from mitochondrial myopathies to cancer [1, 5]. Researchers studying Fe-S cluster biogenesis need reliable tools to investigate the chaperones that mediate cluster delivery. This article provides a comprehensive overview of GO:0140132, covering its definition, mechanism, key genes, disease relevance, and experimental strategies, with a focus on how CRISPR-based models can accelerate discovery.
iron-sulfur cluster chaperone activity At A Glance
| GO ID | GO:0140132 |
|---|---|
| GO term | iron-sulfur cluster chaperone activity |
| Ontology | molecular_function |
| Synonym | iron-sulfur cluster carrier activity |
| Major function | Binding to an iron-sulfur cluster and delivering it to an acceptor molecule |
| Related processes | Iron-sulfur cluster assembly, protein maturation, mitochondrial homeostasis |
| Key chaperones | IscA1, CIAO1, Ssq1p, and other Hsp70-type chaperones |
| Disease relevance | Neuromuscular disorders, cardiomyopathies, cancer |
What Is GO:0140132?
According to the Gene Ontology, GO:0140132 iron-sulfur cluster chaperone activity is a molecular function that involves binding to an iron-sulfur cluster and delivering it to an acceptor molecule [QuickGO]. This activity is also known as iron-sulfur cluster carrier activity. It is distinct from scaffold proteins that assemble clusters; chaperones specifically facilitate the transfer of pre-formed clusters to target apoproteins, ensuring proper metallation and function [2, 8].
Why Is iron-sulfur cluster chaperone activity Important in Cell Biology?
Iron-sulfur cluster chaperone activity is essential for the maturation of Fe-S proteins, which are involved in fundamental cellular processes such as oxidative phosphorylation, DNA repair, and ribosome biogenesis. Dysfunction of these chaperones leads to impaired Fe-S protein activity, resulting in mitochondrial dysfunction and human disease [1, 5]. Therefore, understanding GO:0140132 is crucial for deciphering the molecular basis of Fe-S-related disorders and for developing targeted therapies.
• Fe-S cluster chaperones are required for the activity of enzymes in the electron transport chain, affecting cellular energy production.
• Mutations in the Fe-S cluster chaperone CIAO1 cause a neuromuscular disorder with compromised nucleocytoplasmic Fe-S enzymes.
• Iron-sulfur cluster biogenesis is sensitive to zinc toxicity, highlighting the need for proper chaperone function under stress.
• Sulfur administration can modulate Fe-S cluster homeostasis, implicating chaperones in therapeutic strategies.
• The Hsp70 chaperone Ssq1p is involved in Fe-S cluster formation on the scaffold protein Isu1p, linking chaperone activity to protein folding.
• Chaperone networks are rewired under Coenzyme A and Fe-S cluster deficiency to maintain proteostasis.
• Fe-S cluster chaperones are potential targets for anticancer drugs due to their role in metabolic reprogramming.
• Studying these chaperones can reveal mechanisms of mitochondrial quality control in cardiomyocytes.
• Defects in Fe-S cluster delivery are associated with aging and age-related diseases.
• CRISPR screening can identify novel genes involved in iron-sulfur cluster chaperone activity and their disease connections.
What Happens During iron-sulfur cluster chaperone activity?
Cluster Binding by Chaperones
In simple terms: Chaperones grab the iron-sulfur cluster and hold it safely.
The first step in iron-sulfur cluster chaperone activity is the binding of a pre-formed Fe-S cluster. For example, human IscA1 binds iron with a dissociation constant in the low micromolar range, and this binding is essential for its chaperone function. In mitochondria, the Hsp70 chaperone Ssq1p interacts with the scaffold protein Isu1p to facilitate cluster transfer, although Ssq1p is dispensable for initial cluster formation on Isu1p. This binding protects the cluster from degradation and prepares it for delivery.
Delivery to Acceptor Proteins
In simple terms: The chaperone hands the cluster to the target protein that needs it.
After binding, the chaperone delivers the Fe-S cluster to an acceptor apoprotein. This process often requires specific protein-protein interactions and may be coupled to ATP hydrolysis by Hsp70 chaperones. For instance, CIAO1 in the cytosol is critical for delivering clusters to nucleocytoplasmic Fe-S enzymes, and its loss leads to impaired enzyme activities. The delivery step ensures that the cluster is inserted into the correct target protein, maintaining cellular function.
Regulation by Cellular Stress
In simple terms: When cells are stressed, chaperones change their activity to cope.
Iron-sulfur cluster chaperone activity is regulated in response to cellular stress. Under conditions of Coenzyme A and Fe-S cluster deficiency, the transcription factor HLH-30/TFEB rewires the chaperone network to promote proteostasis. Additionally, zinc toxicity affects Fe-S cluster biogenesis in Escherichia coli, likely by interfering with chaperone function. These regulatory mechanisms ensure that Fe-S protein maturation is prioritized under adverse conditions.
Integration with Mitochondrial Quality Control
In simple terms: Chaperones work with mitochondrial cleanup systems to keep cells healthy.
In cardiomyocytes, Fe-S cluster chaperones are part of the mitochondrial quality control network that safeguards the heart against disease and ageing. This integration ensures that damaged Fe-S proteins are either repaired or degraded, preventing the accumulation of dysfunctional proteins. The chaperone activity is thus tightly linked to mitochondrial proteostasis and overall cellular health.
Key Genes Involved in GO:0140132 iron-sulfur cluster chaperone activity
The following genes encode proteins with iron-sulfur cluster chaperone activity or are directly involved in the delivery of Fe-S clusters to acceptor molecules.
| Gene | Major Role | Research Relevance |
|---|---|---|
| IscA1 | Iron-binding chaperone in mitochondrial Fe-S cluster assembly | Studied for its iron-binding activity and role in cluster transfer |
| CIAO1 | Cytosolic Fe-S cluster chaperone for nucleocytoplasmic enzymes | Loss-of-function causes neuromuscular disorder |
| Ssq1p | Mitochondrial Hsp70 chaperone facilitating cluster transfer to Isu1p | Dispensable for cluster formation but important for transfer |
| Isu1p | Scaffold protein for Fe-S cluster assembly | Interacts with Ssq1p for cluster delivery |
| HLH-30/TFEB | Transcription factor regulating chaperone network under Fe-S deficiency | Rewires chaperone network for proteostasis |
| Hsp70 | General chaperone family involved in protein folding and Fe-S cluster transfer | Potential regulator of Fe-S cluster chaperone activity |
| Grx5 | Mitochondrial monothiol glutaredoxin involved in Fe-S cluster transfer | Not directly cited but part of Fe-S machinery |
| Nfs1 | Cysteine desulfurase providing sulfur for Fe-S clusters | Essential for cluster assembly |
| Frataxin | Mitochondrial protein involved in Fe-S cluster assembly | Defects cause Friedreich ataxia |
| ABCB7 | Mitochondrial transporter involved in Fe-S cluster export | Mutations cause X-linked sideroblastic anemia |
| LYRM4 | Component of the Fe-S cluster assembly complex | Required for cluster assembly |
| NFU1 | Fe-S cluster scaffold/chaperone | Mutations cause multiple mitochondrial dysfunctions syndrome |
| BOLA3 | Fe-S cluster chaperone | Mutations cause multiple mitochondrial dysfunctions syndrome |
| IBA57 | Fe-S cluster chaperone | Mutations cause multiple mitochondrial dysfunctions syndrome |
| CIAO2A | Part of the cytosolic Fe-S cluster assembly machinery | Not directly cited but relevant |
| CIAO2B | Part of the cytosolic Fe-S cluster assembly machinery | Not directly cited but relevant |
| MMS19 | Cytosolic Fe-S cluster assembly factor | Not directly cited but relevant |
How Is iron-sulfur cluster chaperone activity Regulated?
Iron-sulfur cluster chaperone activity is regulated at multiple levels. Under conditions of Coenzyme A and Fe-S cluster deficiency, the transcription factor HLH-30/TFEB rewires the chaperone network to promote proteostasis. Additionally, zinc toxicity can impair Fe-S cluster biogenesis, likely by affecting chaperone function. Sulfur administration has been shown to modulate Fe-S cluster homeostasis, suggesting that sulfur availability regulates chaperone activity. These regulatory mechanisms ensure that Fe-S protein maturation is adapted to cellular stress and metabolic demands.
iron-sulfur cluster chaperone activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CIAO1 | Neuromuscular disorder with compromised nucleocytoplasmic Fe-S enzymes | Knockout or point-mutation in human cell lines (e.g., HEK293) |
| IscA1 | Mitochondrial dysfunction in cardiomyocytes | Cardiomyocyte-specific knockout in mice [1, 2] |
| Ssq1p | Impaired Fe-S cluster transfer in yeast | Yeast knockout and rescue with human homologs |
| HLH-30/TFEB | Proteostasis under Fe-S cluster deficiency | C. elegans knockout and stress assays |
| NFU1 | Multiple mitochondrial dysfunctions syndrome | Patient-derived fibroblasts and CRISPR correction |
Neuromuscular Disorders
Loss-of-function mutations in CIAO1, a cytosolic Fe-S cluster chaperone, cause a neuromuscular disorder characterized by compromised nucleocytoplasmic Fe-S enzymes. This highlights the critical role of chaperone activity in maintaining muscle and nerve function. Patients present with a range of symptoms, including muscle weakness and developmental delay, underscoring the importance of proper Fe-S cluster delivery.
Cardiomyopathies and Ageing
Mitochondrial quality control, including Fe-S cluster chaperone activity, is essential for safeguarding the heart against disease and ageing. Defects in Fe-S cluster assembly lead to mitochondrial dysfunction in cardiomyocytes, contributing to cardiomyopathy and heart failure. Chaperones such as IscA1 help maintain mitochondrial proteostasis, and their impairment accelerates cardiac ageing.
Cancer and Metabolic Reprogramming
Fe-S cluster chaperones are involved in metabolic reprogramming observed in cancer cells. For example, the Hsp70 chaperone Ssq1p and its homologs facilitate cluster transfer to enzymes required for energy metabolism. Targeting these chaperones could disrupt cancer cell metabolism, making them potential therapeutic targets. However, further research is needed to fully elucidate their role in cancer.
Zinc Toxicity and Bacterial Pathogenesis
Zinc toxicity affects Fe-S cluster biogenesis in Escherichia coli, likely by interfering with chaperone function. This suggests that bacterial pathogens may be vulnerable to zinc-based antimicrobial strategies that target Fe-S cluster chaperones. Understanding these mechanisms could lead to new antibiotics.
From iron-sulfur cluster chaperone activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does knockout of CIAO1 impair Fe-S enzyme activity? | CRISPR knockout in HEK293 cells |
| Can a point mutation in IscA1 disrupt iron binding? | CRISPR point mutation in IscA1 |
| Does overexpression of Ssq1p enhance Fe-S cluster transfer? | Yeast overexpression model |
| What is the effect of HLH-30/TFEB knockout on chaperone network? | C. elegans knockout |
| Can knock-in of a tagged CIAO1 reveal its interactome? | CRISPR knock-in of FLAG tag in human cells |
| Does zinc toxicity affect Fe-S cluster chaperone function? | E. coli knockout and zinc treatment |
How to Study the iron-sulfur cluster chaperone activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| UV-visible spectroscopy | Iron-sulfur cluster binding | Assessing IscA1 iron-binding affinity |
| EPR spectroscopy | Paramagnetic Fe-S cluster states | Characterizing cluster transfer intermediates |
| CRISPR knockout screen | Genes required for Fe-S enzyme activity | Identifying novel chaperones |
| Affinity purification-MS | Protein-protein interactions | Mapping CIAO1 interactome |
| RNA-seq | Transcriptional changes under Fe-S deficiency | Studying HLH-30/TFEB regulation |
| Enzyme activity assays | Functional reconstitution of Fe-S enzymes | Measuring cluster transfer to aconitase |
| Yeast genetics | Genetic interactions and complementation | Studying Ssq1p and Isu1p |
| Zinc toxicity assays | Bacterial growth and Fe-S cluster biogenesis | Evaluating zinc effects on chaperones |
Biochemical Assays for Cluster Binding and Transfer
To study iron-sulfur cluster chaperone activity, researchers can use biochemical assays such as iron-binding assays (e.g., UV-visible spectroscopy, EPR) to measure cluster binding to chaperones like IscA1. Cluster transfer to acceptor proteins can be monitored by measuring the reconstitution of enzyme activity (e.g., aconitase) or by using radioactive iron isotopes. These methods provide direct evidence of chaperone function.
CRISPR-Based Genetic Screens
CRISPR knockout screens can identify genes required for Fe-S cluster chaperone activity. For example, a genome-wide screen in human cells could reveal novel chaperones or regulators. Hits can be validated by targeted knockout and biochemical assays. This approach is powerful for discovering new components of the Fe-S cluster delivery pathway.
Proteomics and Interactomics
Proteomic approaches such as affinity purification coupled with mass spectrometry can identify proteins that interact with Fe-S cluster chaperones. For instance, tagging CIAO1 with FLAG and performing pull-downs can reveal its binding partners. Additionally, quantitative proteomics can assess the impact of chaperone loss on Fe-S protein abundance and activity.
Transcriptomics and Stress Response
RNA-seq can be used to study how cells respond to Fe-S cluster deficiency and how chaperone networks are rewired. For example, HLH-30/TFEB target genes can be identified by comparing wild-type and knockout cells under stress. This method helps elucidate regulatory mechanisms controlling chaperone activity.
How CRISPR Can Be Used to Study GO:0140132 iron-sulfur cluster chaperone activity
Knockout
CRISPR knockout of genes encoding Fe-S cluster chaperones (e.g., CIAO1, IscA1) can reveal their essentiality and impact on Fe-S protein maturation. For example, CIAO1 knockout in human cells leads to impaired nucleocytoplasmic Fe-S enzyme activities, mimicking patient phenotypes. Knockout models are valuable for studying loss-of-function effects and for drug screening.
Point Mutation
CRISPR point mutations can be introduced to dissect specific residues required for iron binding or cluster transfer. For instance, mutating iron-coordinating residues in IscA1 can abolish its chaperone activity without affecting protein stability. Such models help distinguish between binding and delivery functions.
Knock-in
Knock-in of epitope tags (e.g., FLAG, HA) into endogenous chaperone genes allows for precise localization and interactome studies. Tagged CIAO1 knock-in cells can be used for affinity purification to identify novel binding partners. This approach maintains physiological expression levels.
Overexpression
CRISPR activation (CRISPRa) or cDNA overexpression can be used to increase chaperone levels and study their effects on Fe-S cluster homeostasis. Overexpression of Ssq1p in yeast can enhance cluster transfer to Isu1p, providing insights into rate-limiting steps. Overexpression models are useful for gain-of-function studies.
How EDITGENE Supports iron-sulfur cluster chaperone activity Research
Researchers studying iron-sulfur cluster chaperone activity-related genes often need to determine whether a candidate gene is causally involved in Fe-S cluster delivery and how its dysfunction contributes to disease. EDITGENE provides a comprehensive suite of CRISPR services to generate precisely engineered cell models, enabling functional validation and mechanistic studies.
Contact EDITGENE today to design your custom CRISPR model for iron-sulfur cluster chaperone activity research.
Frequently Asked Questions About iron-sulfur cluster chaperone activity
What is iron-sulfur cluster chaperone activity?
Iron-sulfur cluster chaperone activity (GO:0140132) is a molecular function where a protein binds to an iron-sulfur cluster and delivers it to an acceptor molecule, ensuring proper maturation of Fe-S proteins [QuickGO].
What genes are involved in iron-sulfur cluster chaperone activity?
Key genes include IscA1, CIAO1, Ssq1p, and HLH-30/TFEB, among others [2, 5, 7, 8].
How is iron-sulfur cluster chaperone activity regulated?
It is regulated by cellular stress pathways, such as the HLH-30/TFEB transcription factor under Fe-S cluster deficiency, and by sulfur availability [4, 7].
What diseases are associated with defects in iron-sulfur cluster chaperones?
Defects are linked to neuromuscular disorders, cardiomyopathies, and cancer [1, 5].
What methods are used to study iron-sulfur cluster chaperone activity?
Common methods include biochemical assays (UV-visible, EPR), CRISPR screens, proteomics, and RNA-seq [2, 5, 7].
Can CRISPR be used to study iron-sulfur cluster chaperone activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are powerful tools to dissect gene function and disease mechanisms [5, 8].
What is the role of CIAO1 in iron-sulfur cluster chaperone activity?
CIAO1 is a cytosolic chaperone that delivers Fe-S clusters to nucleocytoplasmic enzymes; its loss causes a neuromuscular disorder.
How does zinc toxicity affect iron-sulfur cluster biogenesis?
Zinc toxicity impairs Fe-S cluster biogenesis in E. coli, likely by interfering with chaperone function.
What is the connection between iron-sulfur cluster chaperones and mitochondrial quality control?
Chaperones are part of mitochondrial quality control networks that protect cardiomyocytes against disease and ageing.
How can EDITGENE help my research on iron-sulfur cluster chaperone activity?
EDITGENE provides custom CRISPR knockout, point mutation, knock-in, overexpression cell models, and library screening services to study genes involved in Fe-S cluster chaperone activity.
Conclusion
Iron-sulfur cluster chaperone activity (GO:0140132) is a fundamental molecular function required for the maturation of Fe-S proteins, which are critical for cellular metabolism, energy production, and genome maintenance. Dysregulation of this activity leads to severe human diseases, including neuromuscular disorders and cardiomyopathies. Advances in CRISPR-based models and biochemical assays are accelerating our understanding of these chaperones. EDITGENE offers comprehensive services to support researchers in dissecting the mechanisms and disease relevance of iron-sulfur cluster chaperone activity.
References
- 1. Ravindran R et al.. 2025. Mitochondrial quality control in cardiomyocytes: safeguarding the heart against disease and ageing.. Nat Rev Cardiol 22(10):798-813 PMID: 40113864
- 2. Lu J et al.. 2010. Iron-binding activity of human iron-sulfur cluster assembly protein hIscA1.. Biochem J 428(1):125-31 PMID: 20302570
- 3. Li J et al.. 2019. Zinc Toxicity and Iron-Sulfur Cluster Biogenesis in Escherichia coli.. Appl Environ Microbiol 85(9) PMID: 30824435
- 4. Rydz L et al.. 2021. Sulfur Administration in Fe-S Cluster Homeostasis.. Antioxidants (Basel) 10(11) PMID: 34829609
- 5. Maio N et al.. 2024. CIAO1 loss of function causes a neuromuscular disorder with compromise of nucleocytoplasmic Fe-S enzymes.. J Clin Invest 134(12) PMID: 38950322
- 7. Shalash R et al.. 2024. HLH-30/TFEB rewires the chaperone network to promote proteostasis under conditions of Coenzyme A and Iron-Sulfur Cluster Deficiency.. bioRxiv PMID: 38895373
- 8. Dutkiewicz R et al.. 2006. The Hsp70 chaperone Ssq1p is dispensable for iron-sulfur cluster formation on the scaffold protein Isu1p.. J Biol Chem 281(12):7801-8 PMID: 16431909
