Nucleoredoxin, Glutaredoxin, and Thioredoxin Differentially Regulate NF-κB, AP-1, and CREB Activation in HEK293 Cells

doi:10.1006/bbrc.2000.3106

Biochemical and Biophysical Research Communications

Volume 274, Issue 1, 21 July 2000, Pages 177-182

https://doi.org/10.1006/bbrc.2000.3106 Get rights and content

Abstract

Well-established mechanisms for regulation of protein activity include thiol-mediated oxidoreduction in addition to protein–protein interactions and phosphorylation. Nucleoredoxin (NRX), glutaredoxin (GRX), and thioredoxin (TRX) have been shown to act as a potent thiol reductase and reactive oxygen species regulator. They constitute a oxidoreductase superfamily and have been suggested as a candidate operating in the redox regulation of gene expression. We demonstrated here that intracellular localization of these redox molecules differ from each other and that the redox molecules differentially regulate NF-κB, AP-1, and CREB activation induced by TNFα, PMA, and forskolin and by expression of signaling intermediate kinases, NIK, MEKK, and PKA in HEK293 cells. This is a first report that describes involvement of NRX and GRX and differences from TRX in transcriptional regulation of NF-κB, AP-1, and CREB in living cells.

References (27)

A. Holmgren
Thioredoxin and glutaredoxin systems
J. Biol. Chem.
(1989)
H. Kurooka et al.
Cloning and characterization of the nucleoredoxin gene that encodes a novel nuclear protein related to thioredoxin
Genomics
(1997)
Y. Makino et al.
Direct association with thioredoxin allows redox regulation of glucocorticoid receptor function
J. Biol. Chem.
(1999)
Y. Takashima et al.
Differential expression of glutaredoxin and thioredoxin during monocytic differentiation
Immunol. Lett.
(1999)
K. Hirota et al.
Distinct roles of thioredoxin in the cytoplasm and in the nucleus. A two-step mechanism of redox regulation of transcription factor NF-B
J. Biol. Chem
(1999)
M.L. Schmitz et al.
Transactivation domain 2 (TA) of p65 NF-B
J. Biol. Chem.
(1995)
A. Holmgren et al.
Glutaredoxin
Methods Enzymol.
(1995)
F. Åslund et al.
Redox potentials of glutaredoxin and other thiol-disulfide oxidoreductases of the thioredoxin superfamily determined by direct protein–protein redox equilibria
J. Biol. Chem.
(1997)
Y.S. Bae et al.
Epidermal growth factor (EGF)-induced generation of hydrogen peroxide
J. Biol. Chem.
(1997)
A. Mitsui et al.
Reactive oxygen-reducing and protein-refolding activities of adult T cell leukemia-derived factor/human thioredoxin
Biochem. Biophys. Res. Commun.
(1992)

S.W. Kang et al.

Characterization of a mammalian peroxiredoxin that containes one conserved cysteine

J. Biol. Chem.

(1998)

S.W. Kang et al.

Mammalian peroxiredoxin isoforms can reduce hydrogen peroxide generated in response to growth factors and tumor necrosis factor-α

J. Biol. Chem.

(1998)

H.L. Pahl et al.

Oxygen and the control of gene expression

BioEssays

(1994)

Cited by (175)

The significance of glutaredoxins for diabetes mellitus and its complications
2024, Redox Biology
Diabetes mellitus is a non-communicable metabolic disease hallmarked by chronic hyperglycemia caused by beta-cell failure. Diabetic complications affect the vasculature and result in macro- and microangiopathies, which account for a significantly increased morbidity and mortality. The rising incidence and prevalence of diabetes is a major global health burden. There are no feasible strategies for beta-cell preservation available in daily clinical practice. Therefore, patients rely on antidiabetic drugs or the application of exogenous insulin. Glutaredoxins (Grxs) are ubiquitously expressed and highly conserved members of the thioredoxin family of proteins. They have specific functions in redox-mediated signal transduction, iron homeostasis and biosynthesis of iron-sulfur (FeS) proteins, and the regulation of cell proliferation, survival, and function. The involvement of Grxs in chronic diseases has been a topic of research for several decades, suggesting them as therapeutic targets. Little is known about their role in diabetes and its complications. Therefore, this review summarizes the available literature on the significance of Grxs in diabetes and its complications. In conclusion, Grxs are differentially expressed in the endocrine pancreas and in tissues affected by diabetic complications, such as the heart, the kidneys, the eye, and the vasculature. They are involved in several pathways essential for insulin signaling, metabolic inflammation, glucose and fatty acid uptake and processing, cell survival, and iron and mitochondrial metabolism. Most studies describe significant changes in glutaredoxin expression and/or activity in response to the diabetic metabolism.
In general, mitigated levels of Grxs are associated with oxidative distress, cell damage, and even cell death. The induced overexpression is considered a potential part of the cellular stress-response, counteracting oxidative distress and exerting beneficial impact on cell function such as insulin secretion, cytokine expression, and enzyme activity.
Redox Protein Thioredoxin Mediates Neurite Outgrowth in Primary Cultured Mouse Cerebral Cortical Neurons
2024, Neuroscience
Thioredoxin system plays an important role in maintaining the cellular redox balance. Recent evidence suggests that thioredoxin (Trx) system may promote cell survival and neuroprotection. In this study, we explored the role of thioredoxin system in neuronal differentiation using a primary mouse cortical neuronal cell culture. First, Trx and Trx reductase (TrxR) protein levels were analyzed in cultured neurons from 1 to 32 days in vitro (DIV). The result showed that Trx and TrxR protein levels time-dependently increased in the neuron cell culture from 1 to 18 DIV. To establish the role of Trx in neuronal differentiation, Trx gene expression was knockdown in cultured neurons using Trx sgRNA CRISPR/Cas9 technology. Treatment with CRISPR/Cas9/Trx sgRNA decreased Trx protein levels and caused a reduction in dendritic outgrowth and branching of cultured neurons. Then, primary cortical neurons were treated with the Trx inhibitor PX12 to block Trx reducing activity. Treatment with PX12 also reduced dendritic outgrowth and branching. Furthermore, PX12 treatment reduced the ratio of phosphorylated cyclic AMP response element-binding protein (CREB)/total CREB protein levels. To investigate whether CREB phosphorylation is redox regulated, SH-SY5Y cells were treated with H₂O₂, which reduced phosphorylated CREB protein levels and increased CREB thiol oxidation. However, treatment with CB3, a Trx-mimetic tripeptide, rescued H₂O₂-decreased CREB phosphorylation. Our results suggest that Trx regulates neuronal differentiation and maturation of primary mouse cortical neurons by targeting CREB neurotrophic pathway. Trx may regulate CREB activation by maintaining the cellular redox balance.
Impact of selenium nanoparticles in the regulation of inflammation
2022, Archives of Biochemistry and Biophysics
The ability to develop novel medications based on nanoscale complexes has greatly enhanced the capabilities of current pharmaceuticals and has made multidimensional research of these complexes extremely relevant in recent years. Selenium nanoparticles (SeNPs) constitute one such example which in general, could be created by biological, chemical, and physical techniques. Biogenic SeNPs show improved compatibility with human organs and tissues. While sufficient levels of selenium (Se) are crucial for triggering immunity, they also play a role in controlling exaggerated immunological responses and persistent inflammation. More significantly, SeNPs can activate the immune systems, both innate and adaptive, in the tumor microenvironment, which results in an immunological response that fights various diseases caused by chronic inflammation. In this article, we discuss the functions of Se and SeNPs in controlling inflammation with particular emphasis given to their role in combating inflammation in different diseases. Finally, even though Se status exhibits considerable promise as a reliable indicator of autoimmune and inflammatory diseases, novel functionalized SeNPs may likely offer a more effective and reliable tool in both disease prevention and treatment.
Glutaredoxin: Discovery, redox defense and much more
2021, Redox Biology
Glutaredoxin, Grx, is a small protein containing an active site cysteine pair and was discovered in 1976 by Arne Holmgren. The Grx system, comprised of Grx, glutathione, glutathione reductase, and NADPH, was first described as an electron donor for Ribonucleotide Reductase but, from the first discovery in E.coli, the Grx family has impressively grown, particularly in the last two decades. Several isoforms have been described in different organisms (from bacteria to humans) and with different functions.
The unique characteristic of Grxs is their ability to catalyse glutathione-dependent redox regulation via glutathionylation, the conjugation of glutathione to a substrate, and its reverse reaction, deglutathionylation. Grxs have also recently been enrolled in iron sulphur cluster formation. These functions have been implied in various physiological and pathological conditions, from immune defense to neurodegeneration and cancer development thus making Grx a possible drug target.
This review aims to give an overview on Grxs, starting by a phylogenetic analysis of vertebrate Grxs, followed by an analysis of the mechanisms of action, the specific characteristics of the different human isoforms and a discussion on aspects related to human physiology and diseases.
Integrative transcriptome analysis and discovery of genes involving in immune response of hypoxia/thermal challenges in the small abalone Haliotis diversicolor
2019, Fish and Shellfish Immunology
In recent years, the abalone aquaculture industry has been threatened by the deteriorating environmental conditions, such as hypoxia and thermal stress in the hot summers. It is necessary to investigate the molecular mechanism in response to these environmental challenges, and subsequently understand the immune defense system. In this study, the transcriptome profiles by RNA-seq of hemocytes from the small abalone Haliotis diversicolor after exposure to hypoxia, thermal stress, and hypoxia plus thermal stress were established. A total of 103,703,074 clean reads were obtained and 99,774 unigenes were assembled. Of the 99,774 unigenes, 47,154 and 20,455 had homologous sequences in the Nr and Swiss-Prot protein databases, while 16,944 and 10,840 unigenes could be classified by COG or KEGG databases, respectively. RNAseq analysis revealed that the differentially expressed genes (DEGs) after challenges of hypoxia, thermal stress, or hypoxia plus thermal stress were 24,189, 29,165 and 23,665, among which more than 3000 genes involved in at least 230 pathways, including several classical immune-related pathways. The genes and pathways that were involved in immune response to hypoxia/thermal challenges were identified by transcriptome analysis and further validated by quantitative real-time PCR and RNAi technology. The findings in this study can provide information on H. diversicolor innate immunity to improve the abalone aquaculture industry, and the analysis of the potential immune-related genes in innate immunity signaling pathways and the obtained transcriptome data can provide an invaluable genetic resource for the study of the genome and functional genes.
Expression of thioredoxin and glutaredoxin in experimental hepatocellular carcinoma—Relevance for prognostic and diagnostic evaluation
2018, Pathophysiology
Hepatocellular carcinoma (HCC), represents more than 85% of liver cancers. The diagnosis of HCC may be delayed due to the absence of early, sensitive and specific biomarkers. This study was conducted to investigate whether the expression of thioredoxin (Trx) and glutaredoxin (Grx) is helpful for HCC diagnosis in an experimental model. Twenty male albino rats were equally divided into two groups (HCC and control). Hepatocarcinogenesis was performed by single intraperitoneal (i.p) injection of 200 mg/kg of diethylnitrosamine (DENA). Two weeks later, 0.05% of phenobarbital (PB) was supplied in the drinking water for other 14 weeks. HCC was diagnosed by measuring serum alpha-fetoprotein (AFP) level and histopathological examination. Our results found that hepatic indices alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin were elevated but decreased total protein level. Lipid peroxidation was elevated through increasing hepatic content of MDA with decreased antioxidant parameters like hepatic SOD, CAT activities and GSH. The current study also found that Trx and Grx tissue genes were overexpressed in HCC group significantly, compared to control group. This study substantiated that increased expression of these enzymes may be predictive of outcomes in HCC.

View all citing articles on Scopus

^☆: Abbreviations used: NRX; nucleoredoxin, GRX; glutaredoxin; TRX, thioredoxin; NF-κB, nuclear factor κ B; AP-1, activating protein 1; CREB, CRE binding protein; NIK, NF-κB inducible factor; MEKK, MAP/Erk kinase kinase kinase; DMEM; Dulbecco's modified Eagle's medium.

¹: To whom correspondence should be addressed at Department of Anesthesia, Kyoto University Hospital Kyoto University, 54, Shogoin Kawahara-Cho, Sakyo-Ku, Kyoto, 606-8507, Japan. Fax: +81-75-752-3259. E-mail: [email protected].

²: Present address: Laboratory of Biomedical Genetics, Graduate School of Pharmaceutical Sciences, the University of Tokyo, Bunkyo-Ku, Tokyo 113-0033, Japan.

View full text

Regular ArticleNucleoredoxin, Glutaredoxin, and Thioredoxin Differentially Regulate NF-κB, AP-1, and CREB Activation in HEK293 Cells☆

Abstract

J. Biol. Chem.

Genomics

J. Biol. Chem.

Immunol. Lett.

J. Biol. Chem

J. Biol. Chem.

Methods Enzymol.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

J. Biol. Chem.

Oxygen and the control of gene expression

BioEssays

Regular Article
Nucleoredoxin, Glutaredoxin, and Thioredoxin Differentially Regulate NF-κB, AP-1, and CREB Activation in HEK293 Cells☆