MiRNA-210 modulates a nickel-induced cellular energy metabolism shift by repressing the iron–sulfur cluster assembly proteins ISCU1/2 in Neuro-2a cells

ABSTRACTWhile zinc is an essential trace metal in biology, excess zinc is toxic to organisms. Previous studies have shown that zinc toxicity is associated with disruption of the [4Fe-4S] clusters in various dehydratases inEscherichia coli. Here, we report that the intracellular zinc overload inE. colicells inhibits iron-sulfur cluster biogenesis without affecting the preassembled iron-sulfur clusters in proteins. Among the housekeeping iron-sulfur cluster assembly proteins encoded by the gene clusteriscSUA-hscBA-fdx-iscXinE. colicells, the scaffold IscU, the iron chaperone IscA, and ferredoxin have strong zinc binding activity in cells, suggesting that intracellular zinc overload inhibits iron-sulfur cluster biogenesis by binding to the iron-sulfur cluster assembly proteins. Mutations of the conserved cysteine residues to serine in IscA, IscU, or ferredoxin completely abolish the zinc binding activity of the proteins, indicating that zinc can compete with iron or iron-sulfur cluster binding in IscA, IscU, and ferredoxin and block iron-sulfur cluster biogenesis. Furthermore, intracellular zinc overload appears to emulate the slow-growth phenotype of theE. colimutant cells with deletion of the iron-sulfur cluster assembly proteins IscU, IscA, and ferredoxin. Our results suggest that intracellular zinc overload inhibits iron-sulfur cluster biogenesis by targeting the iron-sulfur cluster assembly proteins IscU, IscA, and ferredoxin inE. colicells.IMPORTANCEZinc toxicity has been implicated in causing various human diseases. High concentrations of zinc can also inhibit bacterial cell growth. However, the underlying mechanism has not been fully understood. Here, we report that zinc overload inEscherichia colicells inhibits iron-sulfur cluster biogenesis by targeting specific iron-sulfur cluster assembly proteins. Because iron-sulfur proteins are involved in diverse physiological processes, the zinc-mediated inhibition of iron-sulfur cluster biogenesis could be largely responsible for the zinc-mediated cytotoxicity. Our finding provides new insights on how intracellular zinc overload may inhibit cellular functions in bacteria.

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MicroRNA-210 Modulates the Cellular Energy Metabolism Shift During H2O2-Induced Oxidative Stress by Repressing ISCU in H9c2 Cardiomyocytes

Cellular Physiology and Biochemistry ◽

10.1159/000480417 ◽

2017 ◽

Vol 43 (1) ◽

pp. 383-394 ◽

Cited By ~ 13

Author(s):

Wei Sun ◽

Lei Zhao ◽

Xianjing Song ◽

Jichang Zhang ◽

Yue Xing ◽

...

Keyword(s):

Oxidative Stress ◽

Energy Metabolism ◽

Complex I ◽

Cluster Assembly ◽

Iron Sulfur Cluster ◽

Myocardial Energy Metabolism ◽

Cellular Energy ◽

Atp Assay ◽

Cellular Energy Metabolism ◽

H9c2 Cardiomyocytes

Background/Aims: The myocardial energy metabolism shift is one of the most important pathological features of ischemic heart disease (IHD). Although several microRNAs (miRs) are involved in the regulation of myocardial energy metabolism, their exact effects and underlying mechanisms remain unclear. The aim of this study was to investigate whether microRNA(miR-210) regulates the energy metabolism shift during oxidative stress in H9c2 cardiomyocytes. Methods: Cell survival was analyzed via CCK assay. The energy metabolism shift was detected by lactate assay, ATP assay and RT2 profiler glucose metabolism PCR array. Protein and mRNA expression levels were determined by western blot and qPCR. We also used kits to detect the activity of Complex I, Sirt3 and the NAD+/NADH ratio. Results: We determined that miR-210 promoted the energy metabolism shift. The iron-sulfur cluster assembly protein (ISCU) was a target of miR-210. Additionally, we detected the activity of complex I and found that miR-210 inhibits mitochondrial respiration. Interestingly, miR-210 may also indirectly regulate SIRT3 by regulating ISCU. Conclusion: Our results confirm that miR-210 is essential and sufficient for modulating the cellular energy metabolism shift during H2O2-induced oxidative stress in H9c2 cardiomyocytes by targeting ISCU.

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MiRNA-210 modulates nickel-induced hypoxic responses by repressing the iron-sulfur cluster assembly proteins ISCU1/2

Toxicology Letters ◽

10.1016/j.toxlet.2013.05.320 ◽

2013 ◽

Vol 221 ◽

pp. S154

Author(s):

Mindi He ◽

Lin Mao ◽

Zhengping Yu ◽

Zhou Zhou

Keyword(s):

Cluster Assembly ◽

Iron Sulfur Cluster ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Assembly Proteins

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Knock-downs of Iron-Sulfur Cluster Assembly Proteins IscS and IscU Down-regulate the Active Mitochondrion of ProcyclicTrypanosoma brucei

Journal of Biological Chemistry ◽

10.1074/jbc.m513781200 ◽

2006 ◽

Vol 281 (39) ◽

pp. 28679-28686 ◽

Cited By ~ 53

Author(s):

Ondrej Smíd ◽

Eva Horáková ◽

Vanda Vilímová ◽

Ivan Hrdý ◽

Richard Cammack ◽

...

Keyword(s):

Cluster Assembly ◽

Iron Sulfur Cluster ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Assembly Proteins

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Network of Protein-Protein Interactions among Iron-Sulfur Cluster Assembly Proteins in Escherichia coli1

The Journal of Biochemistry ◽

10.1093/oxfordjournals.jbchem.a003156 ◽

2002 ◽

Vol 131 (5) ◽

pp. 713-719 ◽

Cited By ~ 77

Author(s):

U. Tokumoto ◽

S. Nomura ◽

Y. Minami ◽

H. Mihara ◽

S.-i. Kato ◽

...

Keyword(s):

Protein Interactions ◽

Cluster Assembly ◽

Protein Protein Interactions ◽

Iron Sulfur Cluster ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Assembly Proteins

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Effect of iron–sulfur cluster assembly proteins on the expression of Escherichia coli lipoic acid synthase

Protein Expression and Purification ◽

10.1016/s1046-5928(02)00680-0 ◽

2003 ◽

Vol 28 (2) ◽

pp. 241-245 ◽

Cited By ~ 22

Author(s):

Marco Kriek ◽

Liv Peters ◽

Yasahiro Takahashi ◽

Peter L. Roach

Keyword(s):

Escherichia Coli ◽

Lipoic Acid ◽

Cluster Assembly ◽

Iron Sulfur Cluster ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Assembly Proteins

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MicroRNA-210 Controls Mitochondrial Metabolism during Hypoxia by Repressing the Iron-Sulfur Cluster Assembly Proteins ISCU1/2

Cell Metabolism ◽

10.1016/j.cmet.2009.08.015 ◽

2009 ◽

Vol 10 (4) ◽

pp. 273-284 ◽

Cited By ~ 395

Author(s):

Stephen Y. Chan ◽

Ying-Yi Zhang ◽

Craig Hemann ◽

Christopher E. Mahoney ◽

Jay L. Zweier ◽

...

Keyword(s):

Mitochondrial Metabolism ◽

Cluster Assembly ◽

Iron Sulfur Cluster ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Assembly Proteins

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A novel role of the mitochondrial iron-sulfur cluster assembly protein ISCU-1/ISCU in longevity and stress response

GeroScience ◽

10.1007/s11357-021-00327-z ◽

2021 ◽

Author(s):

Yi Sheng ◽

Guang Yang ◽

Kaitlyn Casey ◽

Shayla Curry ◽

Mason Oliver ◽

...

Keyword(s):

Stress Response ◽

Cluster Assembly ◽

Iron Sulfur Cluster ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Mitochondrial Iron

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Multiple Sense and Antisense Promoters Contribute to the Regulated Expression of the isc-suf Operon for Iron-Sulfur Cluster Assembly in Rhodobacter

Microorganisms ◽

10.3390/microorganisms7120671 ◽

2019 ◽

Vol 7 (12) ◽

pp. 671 ◽

Cited By ~ 1

Author(s):

Xin Nie ◽

Bernhard Remes ◽

Gabriele Klug

Keyword(s):

Rhodobacter Sphaeroides ◽

Photosynthetic Electron Transport ◽

Regulatory Proteins ◽

Cluster Assembly ◽

Iron Sulfur Cluster ◽

Iron Sulfur Clusters ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Photosynthetic Complexes ◽

The One

A multitude of biological functions relies on iron-sulfur clusters. The formation of photosynthetic complexes goes along with an additional demand for iron-sulfur clusters for bacteriochlorophyll synthesis and photosynthetic electron transport. However, photooxidative stress leads to the destruction of iron-sulfur clusters, and the released iron promotes the formation of further reactive oxygen species. A balanced regulation of iron-sulfur cluster synthesis is required to guarantee the supply of this cofactor, on the one hand, but also to limit stress, on the other hand. The phototrophic alpha-proteobacterium Rhodobacter sphaeroides harbors a large operon for iron-sulfur cluster assembly comprising the iscRS and suf genes. IscR (iron-sulfur cluster regulator) is an iron-dependent regulator of isc-suf genes and other genes with a role in iron metabolism. We applied reporter gene fusions to identify promoters of the isc-suf operon and studied their activity alone or in combination under different conditions. Gel-retardation assays showed the binding of regulatory proteins to individual promoters. Our results demonstrated that several promoters in a sense and antisense direction influenced isc-suf expression and the binding of the IscR, Irr, and OxyR regulatory proteins to individual promoters. These findings demonstrated a complex regulatory network of several promoters and regulatory proteins that helped to adjust iron-sulfur cluster assembly to changing conditions in Rhodobacter sphaeroides.

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