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Association of casein kinase II with NF-B occurs in the cytosolic compartment. a, nuclear and cytoplasmic extracts were made from HepG2 cells after treatment for increasing times with 20 ng/ml IL-1 as indicated. NF-B was immunoprecipitated using anti-p65 antibody. The washed immunoprecipitates were assayed for associated-and-casein kinase activities in the presence (bottom panels) or absence (top panels) of heparin. Similar results were obtained in two independent experiments. b, aliquots of the nuclear and cytoplasmic extracts shown in panel a were separated by SDS-PAGE, transferred to nitrocellulose, and immunoblotted with anti-NF-B p65 antibody. Immune complexes were visualized using an enhanced chemiluminescence (ECL, Amersham Corp.) procedure. c, quantitative analysis of the data for-casein phosphorylation presented in panel a. Quantitation of-casein phosphorylation gave essentially identical results but has been omitted for clarity.
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In fibroblasts and hepatoma cells, interleukin-1 (IL-1) treatment results in the rapid nuclear accumulation of the transcription factor NF-kappaB, present largely as p65 (RelA)/p50 heterodimers. It is well established that this process is dependent in large part upon the phosphorylation and subsequent degradation of the cytosolic inhibitor IkappaB....
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... from casein kinase II and that their association occurs simply as a consequence of nuclear translocation of the transcription factor. Accordingly, cytosolic and nuclear extracts were prepared from HepG2 cells after various periods of IL-1 stimulation and immunoprecipitated with anti-p65 for determination of associated casein kinase II activity (Fig. 6, a and c). A casein kinase II-associated pool of NF-B is established first in the cytoplasm and is detectable between 2 and 5 min after addition of IL-1. Accumulation of this form of NF-B in the nucleus is delayed until the 15 min time point is reached, and peak levels of casein kinase II activity associated with both nuclear and cytoplasmic ...
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... until the 15 min time point is reached, and peak levels of casein kinase II activity associated with both nuclear and cytoplasmic NF-B are reached after 30 min of IL-1 treatment. The level of casein kinase II detectable in the nuclear fraction correlates well with the appearance of immunoreactive p65 in the nucleus, as shown by Western blotting (Fig. 6b). These data are consistent with a model in which casein kinase II associates with (and phosphorylates) cytoplasmic NF-B p65 which then translo- cates to the nucleus. An obvious mechanism, although difficult to directly test at the present time, would be one in which dissociation of IB from p50/p65 heterodimers exposes a phos- ...
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... Among the deregulated proteins, the top five proteins with the highest fold change expression values were the following: casein kinase 2 alpha 1 (CSNK2A1), A-Raf proto-oncogene serine/threonine kinase (ARAF), mitogen-activated protein kinase kinase kinase kinase 4 (MAP4K4), phosphoinositide-3-kinase regulatory subunit 2 (PIK3R2), and tyrosyl-DNA phosphodiesterase 2 (TDP2). CSNK2A1, during TNF-α and IL-1 receptors activation, phosphorylates the p65 (RelA) NF-κB subunit in the cytoplasm, leading to its translocation to the nucleus [66][67][68][69][70]. A-Raf activates MEKK1 in the cytoplasm, leading to NF-κB activation [71]. ...
HIV-associated neurocognitive disorders (HAND) affect 15–55% of HIV-positive patients and effective therapies are unavailable. HIV-infected monocyte-derived macrophages (MDM) invade the brain of these individuals, promoting neurotoxicity. We demonstrated an increased expression of cathepsin B (CATB), a lysosomal protease, in monocytes and post-mortem brain tissues of women with HAND. Increased CATB release from HIV-infected MDM leads to neurotoxicity, and their secretion is associated with NF-κB activation, oxidative stress, and lysosomal exocytosis. Cannabinoid receptor 2 (CB2R) agonist, JWH-133, decreases HIV-1 replication, CATB secretion, and neurotoxicity from HIV-infected MDM, but the mechanisms are not entirely understood. We hypothesized that HIV-1 infection upregulates the expression of proteins associated with oxidative stress and that a CB2R agonist could reverse these effects. MDM were isolated from healthy women donors (n = 3), infected with HIV-1ADA, and treated with JWH-133. After 13 days post-infection, cell lysates were labeled by Tandem Mass Tag (TMT) and analyzed by LC/MS/MS quantitative proteomics bioinformatics. While HIV-1 infection upregulated CATB, NF-κB signaling, Nrf2-mediated oxidative stress response, and lysosomal exocytosis, JWH-133 treatment downregulated the expression of the proteins involved in these pathways. Our results suggest that JWH-133 is a potential alternative therapy against HIV-induced neurotoxicity and warrant in vivo studies to test its potential against HAND.
... In line with our findings, a previous study reported that TNF-α stimulates nuclear translocation of p65 in human cancer cells [68,69]. Accordingly, IL-1β is linked to a rapid nuclear accumulation of p65 in human hepatoma cells [70]. Multiple studies have already demonstrated the influence of sex on the immunological response. ...
Sex-related differences are a current topic in contemporary science. In addition to hormonal regulation, cell-autonomous mechanisms are important in bone homeostasis and regeneration. In this study, human skeletal stem cells (SSCs) from female and male adults were cultured and analyzed with immunological assays and osteogenic differentiation assessments. Female SSCs exhibited a mean doubling time of 100.6 h, whereas male SSCs displayed a mean doubling time of 168.0 h. Immunophenotyping revealed the expression of the stem cell markers Nestin, CD133, and CD164, accompanied by the neural-crest marker SOX9. Furthermore, multiparameter flow cytometric analyses revealed a substantial population of multipotent SSCs, comprising up to 80% in both sexes. An analysis of the osteogenic differentiation potential demonstrated a strong mineralization in both male and female SSCs under physiological conditions. Recognizing the prevailing association of bone diseases with inflammatory processes, we also analyzed the osteogenic potential of SSCs from both sexes under pro-inflammatory conditions. Upon TNF-α and IL-1β treatment, we observed no sexual dimorphism on osteogenesis. In summary, we demonstrated the successful isolation and characterization of SSCs capable of rapid osteogenic differentiation. Taken together, in vitro cultured SSCs might be a suitable model to study sexual dimorphisms and develop drugs for degenerative bone diseases.
... CDDO-Me directly or indirectly inhibits the NF-κB pathway [38]. Furthermore, CDDO-Me inhibits casein kinase 2 (CK2) that regulates NF-κB activity [11,39]. Since CK2 enhances NF-κB nuclear transcriptional activity by S529 phosphorylation [40,41], our findings indicate that CDDO-Me may also ameliorate clasmatodendrosis by inhibiting CK2-mediated NF-κB S529 phosphorylation. ...
... In the present study, we found that both CDDO-Me and SN50 ameliorated aberrant mitochondrial hyperfusion by recovering NF-κB-PDI-mediated S-nitrosylation of DRP1. Interestingly, CK2 is an upstream regulator of both NF-κB S529 phosphorylation and AKT activation during clasmatodendrosis [11,[39][40][41]. Considering oxidative stress-induced CK2 activation [11] and the anti-oxidant properties of CDDO-Me [24,25], our findings suggest that oxidative stress may cause CK2 hyperactivation eliciting clasmatodendrosis through the NF-κB-PDIand AKT-Bif-1-mediated signaling pathways. ...
Clasmatodendrosis is a kind of astroglial degeneration pattern which facilitates excessive autophagy. Although abnormal mitochondrial elongation is relevant to this astroglial degeneration, the underlying mechanisms of aberrant mitochondrial dynamics are still incompletely understood. Protein disulfide isomerase (PDI) is an oxidoreductase in the endoplasmic reticulum (ER). Since PDI expression is downregulated in clasmatodendritic astrocytes, PDI may be involved in aberrant mitochondrial elongation in clasmatodendritic astrocytes. In the present study, 26% of CA1 astrocytes showed clasmatodendritic degeneration in chronic epilepsy rats. 2-cyano-3,12-dioxo-oleana-1,9(11)-dien-28-oic acid methyl ester (CDDO-Me; bardoxolone methyl or RTA 402) and SN50 (a nuclear factor-κB (NF-κB) inhibitor) ameliorated the fraction of clasmatodendritic astrocytes to 6.8 and 8.1% in CA1 astrocytes, accompanied by the decreases in lysosomal-associated membrane protein 1 (LAMP1) expression and microtubule-associated protein 1A/1B light-chain 3 (LC3)-II/LC3-I ratio, indicating the reduced autophagy flux. Furthermore, CDDO-Me and SN50 reduced NF-κB S529 fluorescent intensity to 0.6- and 0.57-fold of vehicle-treated animal level, respectively. CDDO-Me and SN50 facilitated mitochondrial fission in CA1 astrocytes, independent of dynamin-related protein 1 (DRP1) S616 phosphorylation. In chronic epilepsy rats, total PDI protein, S-nitrosylated PDI (SNO-PDI), and SNO-DRP1 levels were 0.35-, 0.34- and 0.45-fold of control level, respectively, in the CA1 region and increased CDDO-Me and SN50. Furthermore, PDI knockdown resulted in mitochondrial elongation in intact CA1 astrocytes under physiological condition, while it did not evoke clasmatodendrosis. Therefore, our findings suggest that NF-κB-mediated PDI inhibition may play an important role in clasmatodendrosis via aberrant mitochondrial elongation.
... The ability to interact with all DNA-dependent RNA polymerases has suggested a role for CK2 in gene regulation [120]. Among CK2 targets, we recall here the RPB1 subunit of RNA polymerase II [121], several transcription factors [122][123][124][125][126][127], various components of the spliceosome [128][129][130] and its regulators [121,131], as well as some components of the chromatin, thus influencing its remodeling (reviewed in [118]). As such, CK2 is one of the most studied kinases as a promising target for anti-cancer drug development [132]. ...
Cytokinesis, the conclusive act of cell division, allows cytoplasmic organelles and chromosomes to be faithfully partitioned between two daughter cells. In animal organisms, its accurate regulation is a fundamental task for normal development and for preventing aneuploidy. Cytokinesis failures produce genetically unstable tetraploid cells and ultimately result in chromosome instability, a hallmark of cancer cells. In animal cells, the assembly and constriction of an actomyosin ring drive cleavage furrow ingression, resulting in the formation of a cytoplasmic intercellular bridge, which is severed during abscission, the final event of cytokinesis. Kinase-mediated phosphorylation is a crucial process to orchestrate the spatio-temporal regulation of the different stages of cytokinesis. Several kinases have been described in the literature, such as cyclin-dependent kinase, polo-like kinase 1, and Aurora B, regulating both furrow ingression and/or abscission. However, others exist, with well-established roles in cell-cycle progression but whose specific role in cytokinesis has been poorly investigated, leading to considering these kinases as “minor” actors in this process. Yet, they deserve additional attention, as they might disclose unexpected routes of cell division regulation. Here, we summarize the role of multifunctional kinases in cytokinesis with a special focus on those with a still scarcely defined function during cell cleavage. Moreover, we discuss their implication in cancer.
... Relating to gene expression, CK2 interacts with and phosphorylates a variety of transcription factors (see, e.g., [101][102][103]). The effects of these events range from altered localization, stability, activation state, or association with other binding partner molecules. ...
The association of protein kinase CK2 (formerly casein kinase II or 2) with cell growth and proliferation in cells was apparent at early stages of its investigation. A cancer-specific role for CK2 remained unclear until it was determined that CK2 was also a potent suppressor of cell death (apoptosis); the latter characteristic differentiated its function in normal versus malignant cells because dysregulation of both cell growth and cell death is a universal feature of cancer cells. Over time, it became evident that CK2 exerts its influence on a diverse range of cell functions in normal as well as in transformed cells. As such, CK2 and its substrates are localized in various compartments of the cell. The dysregulation of CK2 is documented in a wide range of malignancies; notably, by increased CK2 protein and activity levels with relatively moderate change in its RNA abundance. High levels of CK2 are associated with poor prognosis in multiple cancer types, and CK2 is a target for active research and testing for cancer therapy. Aspects of CK2 cellular roles and targeting in cancer are discussed in the present review, with focus on nuclear and mitochondrial functions and prostate, breast and head and neck malignancies.
... With a collection of more than 300 known substrates, it is not surprising that several inflammatory mediators are phosphorylated by CK2. NF-κB, STAT1, STAT3, CREB, CREM, Sp1, and C/EBP are examples of transcription factors regulated by CK2 with a role in inflammation and associated disorders (Bird et al., 1997;Yamaguchi et al., 1998;Filhol et al., 2004;Singh and Ramji, 2008;Drygin et al., 2011). NF-κB regulation by CK2 has been extensively characterized in epithelial, HeLa, and diploid gingival fibroblast cells (Bird et al., 1997;Wang et al., 2000;Parhar et al., 2007;Singh and Ramji, 2008). ...
... NF-κB, STAT1, STAT3, CREB, CREM, Sp1, and C/EBP are examples of transcription factors regulated by CK2 with a role in inflammation and associated disorders (Bird et al., 1997;Yamaguchi et al., 1998;Filhol et al., 2004;Singh and Ramji, 2008;Drygin et al., 2011). NF-κB regulation by CK2 has been extensively characterized in epithelial, HeLa, and diploid gingival fibroblast cells (Bird et al., 1997;Wang et al., 2000;Parhar et al., 2007;Singh and Ramji, 2008). Furthermore, CK2 activity is stimulated following treatment with proinflammatory agents (LPS) or cytokines (TNF-α, IL-1, TGF-β, or IFN-γ) in a variety of murine and human cell types (Lodie et al., 1997;Rosenberger et al., 2016). ...
Novel treatments for neurodegenerative disorders are in high demand. It is imperative that new protein targets be identified to address this need. Characterization and validation of nascent targets can be accomplished very effectively using highly specific and potent chemical probes. Human induced pluripotent stem cells (hiPSCs) provide a relevant platform for testing new compounds in disease relevant cell types. However, many recent studies utilizing this platform have focused on neuronal cells. In this study, we used hiPSC-derived microglia-like cells (MGLs) to perform side-by-side testing of a selective chemical probe, SGC-CK2-1, compared with an advanced clinical candidate, CX-4945, both targeting casein kinase 2 (CK2), one of the first kinases shown to be dysregulated in Alzheimer’s disease (AD). CK2 can mediate neuroinflammation in AD, however, its role in microglia, the innate immune cells of the central nervous system (CNS), has not been defined. We analyzed available RNA-seq data to determine the microglial expression of kinases inhibited by SGC-CK2-1 and CX-4945 with a reported role in mediating inflammation in glial cells. As proof-of-concept for using hiPSC-MGLs as a potential screening platform, we used both wild-type (WT) MGLs and MGLs harboring a mutation in presenilin-1 (PSEN1), which is causative for early-onset, familial AD (FAD). We stimulated these MGLs with pro-inflammatory lipopolysaccharides (LPS) derived from E. coli and observed strong inhibition of the expression and secretion of proinflammatory cytokines by simultaneous treatment with SGC-CK2-1. A direct comparison shows that SGC-CK2-1 was more effective at suppression of proinflammatory cytokines than CX-4945. Together, these results validate a selective chemical probe, SGC-CK2-1, in human microglia as a tool to reduce neuroinflammation.
... It is a consensus that the phosphorylation of p65 is a necessary condition for nuclear localization of NF-kappaB to play a cancer-promoting effect (Viatour et al., 2005). Therefore, OTUB2 may be involved in the K48-mediated degradation of proteins that are responsible for the phosphorylation of p65, such as casein kinase 2 (CK2) (Bird et al., 1997;Wang et al., 2000), eventually, OTUB2 perhaps indirectly lead to the reduction of phosphorylation modification of p65. Of course, this approach is worth exploring with experiments. ...
Ovarian tumor (OTU) domain-containing ubiquitin aldehyde-binding protein Otubain2 (OTUB2) was a functional cysteine protease in the OTU family with deubiquitinase activity. In recent years, with the wide application of molecular biology techniques, molecular mechanism regulation at multiple levels of cell signaling pathways has been gradually known, such as ubiquitin-mediated protein degradation and phosphorylation-mediated protein activation. OTUB2 is involved in the deubiquitination of many key proteins in different cell signaling pathways, and the effect of OTUB2 on human health or disease is not clear. OTUB2 is likely to cause cancer and other malignant diseases while maintaining normal human development and physiological function. Therefore, it is of great value to comprehensively understand the regulatory mechanism of OTUB2 and regard it as a target for the treatment of diseases. This review makes a general description and appropriate analysis of OTUB2’s regulation in different cell signaling pathways, and connects OTUB2 with cancer from the research hotspot perspective of DNA damage repair and immunity, laying the theoretical foundation for future research.
... 81,85,86 Although metastatic potential, cell proliferation, apoptotic evasion, and acute inflammatory response have been linked to both pathway arms, the noncanonical pathway may have a more important role in prolonged inflammation owing to its delayed activation through CD40 ligand (CD40L), TNF superfamily member 13b (TNSF13B or BAFF), and TNF superfamily member 12 (TNFS12 or TWEAK) ligands, among others. 84,85,[87][88][89] Canonical activation on the other hand can be induced by classic ligand-receptor cytokines such as IL1b, IL8, and TNFa, as defined earlier [90][91][92][93] ( Figure 3). In addition, NF-kB is capable of transcribing pro-IL1b, IL8, and TNFa. ...
There is no doubt that chronic gastro-esophageal reflux disease (GERD) increases the risk of esophageal adenocarcinoma (EAC) by several fold (OR=6.4, CI=4.6 to 9.1), and some relationships between reflux disease-mediated inflammation and oncogenic processes have been explored; however, the precise interconnections between the immune response and genomic instabilities underlying these pathological processes are only now emerging. Furthermore, the precise cell of origin of the pre-cancerous stages associated with EAC development, Barrett’s esophagus (BE), be it cardia resident or embryonic remnant, may shape our interpretation of the likely immune drivers. This review integrates the current collective knowledge of the immunology underlying EAC development and outlines a framework connecting pro-inflammatory pathways— such as those mediated by IL1β, TNFα, LIF, IL6, STAT3, NF-κB, COX2 and TGFβ—with oncogenic pathways in the GERD-BE-EAC cancer sequence. Further defining these immune and molecular “railroads” may reveal a map of the routes taken by gastro-esophageal cells on their journey toward EAC tumour phylogeny. The selective pressures, applied by this immune-induced journey, likely impact the phenotype and genotype of the resulting oncogenic destination and further exploration of lesser defined immune drivers may be useful in future individualized therapies or enhanced selective application of recent immune-driven therapeutics.
... Squamous cell carcinoma of the head and neck treated with interleukin-6 for 30 min shows an increment in total CK2 activity (Su et al. 2011). Fibroblast and hepatoma cells treated with interleukin-1 show an increase in CK2 activity that is responsible for phosphorylation and nuclear translocation of NF-KB p65 (Bird et al. 1997). HEK293 cells treated with a mixture of inflammatory cytokines plus aldosterone show an increase in protein expression of CK2a and CK2b but not CK2a' while the mRNA level of all CK2 subunits was not affected (Ruhs et al. 2017). ...
CK2 is a constitutively active protein kinase that assuring a constant level of phosphorylation to its numerous substrates supports many of the most important biological functions. Nevertheless, its activity has to be controlled and adjusted in order to cope with the varying needs of a cell, and several examples of a fine-tune regulation of its activity have been described. More importantly, aberrant regulation of this enzyme may have pathological consequences, e.g. in cancer, chronic inflammation, neurodegeneration, and viral infection. Our review aims at summarizing our current knowledge about CK2 regulation. In the first part, we have considered the most important stimuli shown to affect protein kinase CK2 activity/expression. In the second part, we focus on the molecular mechanisms by which CK2 can be regulated, discussing controversial aspects and future perspectives.
... NF-κB is a critical mediator of LPS-induced inflammation (Gloire et al., 2006). NF-κB triggers various pro-inflammatory genes, including TNF-α, which also activates NF-κB, generating a positive feedback loop and leading to a cytokine cascade reaction and the amplification of inflammation (Baeuerle and Henkel, 1994;Baldwin, 1996;Bird et al., 1997). The activation of NF-κB is tightly controlled by the IKK complex and its substrate IκBα. ...
Purpose: Adiponectin has been shown to exert potent anti-inflammatory activities in a range of systemic inflammatory diseases. This study aimed to investigate the potential therapeutic effects of KS23, a globular adiponectin-derived peptide, on endotoxin-induced uveitis (EIU) in rats and lipopolysaccharide (LPS)-stimulated mouse macrophage-like RAW 264.7 cells.
Methods: EIU was induced in Lewis rats by subcutaneous injection of LPS into a single footpad. KS23 or phosphate-buffered saline (PBS) was administered immediately after LPS induction via intravitreal injection. Twenty-four hours later, clinical and histopathological scores were evaluated, and the aqueous humor (AqH) was collected to determine the infiltrating cells, protein concentration, and levels of inflammatory cytokines. In vitro, cultured RAW 264.7 cells were stimulated with LPS in the presence or absence of KS23, inflammatory cytokine levels in the supernatant, nuclear translocation of nuclear factor kappa B (NF-κB) subunit p65, and the expression of NF-kB signaling pathway components were analyzed.
Results: KS23 treatment significantly ameliorated the clinical and histopathological scores of EIU rats and reduced the levels of infiltration cells, protein, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the aqueous humor. Consistently, KS23 decreased the expression of TNF-α and IL-6 in the supernatant of LPS-stimulated RAW 264.7 cells and inhibited the LPS-induced nuclear translocation of NF-κB p65 and the phosphorylation of IKKα/β/IκBα/NF-κB.
Conclusion: The in vivo and in vitro results demonstrated the anti-inflammatory effects of the peptide KS23 and suggested that KS23 is a compelling, novel therapeutic candidate for the treatment of ocular inflammation.
