Evidence that interleukin-1 and phorbol esters activate NF-kappa B by different pathways: role of protein kinase C.

Nuclear factor kappa B (NF-kappa B) is a ubiquitous transcription factor that affects expression of many genes, including immunoglobulin kappa (kappa), the interleukin-2 receptor alpha chain, and two genes in HIV-1. NF-kappa B can be activated by a number of stimuli, including pharmacological stimulation of protein kinase C by phorbol 12-myristate 13-acetate (PMA) and treatment in vitro with either protein kinase C or protein kinase A. This has lead to the proposal that these kinases are key enzymes in the physiological activation of NF-kappa B as well. We have used a murine B cell line, 70Z/3, and T cell line, EL-4 6.1 C10, to study the activation of NF-kappa B by two physiological activators, interleukin-1 alpha (IL-1) and lipopolysaccharide (LPS). There are four reasons to propose that these agents activate pathways that do not include protein kinase C as a major component in these cell lines. First, the protein kinase C inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) strongly inhibited PMA-induced activation of NF-kappa B in 70Z/3 cells but had no effect on NF-kappa B activated by IL-1 or LPS. Second, depletion of protein kinase C by prolonged growth of 70Z/3 in PMA abrogated the capacity of the cells to activate NF-kappa B in response to further PMA treatment. However, these same cells activated NF-kappa B normally after either IL-1 or LPS treatment. Third, IL-1 effectively activated NF-kappa B in EL-4 6.1 C10 cells, but PMA did not. Fourth, interferon-gamma is a potent activator of protein kinase C in 70Z/3 cells, but is completely inactive in the mobilization of NF-kappa B. These results suggest that the physiological inducers IL-1 and LPS activate NF-kappa B by pathways independent of protein kinase C in both 70Z/3 and EL-4 6.1 C10 cells.

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Contrasting effects of the protein kinase C inhibitor staurosporine on the interleukin-1 and phorbol ester activation pathways in the EL4-6.1 thymoma cell line

Journal of Cellular Physiology ◽

10.1002/jcp.1041510112 ◽

1992 ◽

Vol 151 (1) ◽

pp. 71-80 ◽

Cited By ~ 20

Author(s):

Jacques Dornand ◽

Monsif Bouaboula ◽

Arnaud Dupuy D'Angeac ◽

Jean Favero ◽

David Shire ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Cell Line ◽

Phorbol Ester ◽

Interleukin 1 ◽

Kinase C ◽

Protein Kinase C Inhibitor ◽

Activation Pathways

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A role for protein kinase C activity in interleukin-1 (IL-1) induction of IL-2 gene expression but not in IL-1 signal transduction.

Molecular and Cellular Biology ◽

10.1128/mcb.10.6.2731 ◽

1990 ◽

Vol 10 (6) ◽

pp. 2731-2737 ◽

Cited By ~ 19

Author(s):

G Macchia ◽

C T Baldari ◽

A Massone ◽

J L Telford

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Phorbol Myristate Acetate ◽

Phorbol Esters ◽

Interleukin 1 ◽

Interleukin 2 ◽

Calcium Ionophore ◽

Myristate Acetate ◽

Bacterial Gene ◽

Kinase C

Interleukin-1 (IL-1) is known to synergize with phorbol esters in the induction of interleukin-2 (IL-2) expression in T-lymphoid leukemia cells and proliferation of mouse thymocytes. We used a plasmid construct containing the bacterial gene for chloramphenicol acetyltransferase under the control of the human IL-2 promoter to study the nature of this synergism in the murine thymoma cell line EL4. Although IL-1 induction of the IL-2 promoter in these cells required costimulus with phorbol myristate acetate, the signal induced by IL-1 was qualitatively different. We provide evidence to support the hypothesis that the phorbol ester signal is mediated by protein kinase C, and we show that the IL-1 signal is not. That IL-1 and phorbol myristate acetate represent different stimuli was shown by their response to protein kinase C inhibitors, capacity to synergize with increased intracellular free calcium, and requirement for protein synthesis. In addition we show that pretreatment with IL-1 can prime EL4 cells to subsequent activation by concentrations of phorbol esters not normally sufficient to induce IL-2 expression. Pretreated cells remained primed for at least 40 h after removal of the IL-1. Neither phorbol myristate acetate nor a calcium ionophore was capable of preactivating EL4 cells.

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A role for protein kinase C activity in interleukin-1 (IL-1) induction of IL-2 gene expression but not in IL-1 signal transduction

Molecular and Cellular Biology ◽

10.1128/mcb.10.6.2731-2737.1990 ◽

1990 ◽

Vol 10 (6) ◽

pp. 2731-2737

Author(s):

G Macchia ◽

C T Baldari ◽

A Massone ◽

J L Telford

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Phorbol Myristate Acetate ◽

Phorbol Esters ◽

Interleukin 1 ◽

Interleukin 2 ◽

Calcium Ionophore ◽

Myristate Acetate ◽

Bacterial Gene ◽

Kinase C

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Regulation of protein kinase C activity in neuronal differentiation induced by the N-ras oncogene in PC-12 cells

Molecular and Cellular Biology ◽

10.1128/mcb.10.6.2983-2990.1990 ◽

1990 ◽

Vol 10 (6) ◽

pp. 2983-2990

Author(s):

J C Lacal ◽

A Cuadrado ◽

J E Jones ◽

R Trotta ◽

D E Burstein ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Neuronal Differentiation ◽

Phorbol Esters ◽

Ras Oncogene ◽

Intact Cells ◽

Pc 12 Cells ◽

Kinase C ◽

Ras P21

Expression of the N-ras oncogene under the control of the glucocorticoid-responsive promoter in the pheochromocytoma cell line UR61, a subline of PC-12 cells, has been used to investigate the differentiation process to neuronal cells triggered by ras oncogenes (I. Guerrero, A. Pellicer, and D. E. Burstein, Biochem. Biophys. Res. Commun. 150:1185-1192, 1988). Using ras-inducible cell lines, we observed that expression of the oncogenic N-ras p21 protein interferes with the ability of phorbol esters to induce downregulation of protein kinase C. This effect was associated with the appearance of immunologically detectable protein kinase C as well as the activity of the enzyme as analyzed either by binding of [3H]phorbol-12,13-dibutyrate in intact cells or by in vitro kinase activity. These results indicate a relationship between ras p21 and protein kinase C in neuronal differentiation in this model system. Comparison to the murine fibroblast system suggests that this relationship may be functional.

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Formulation of Liver-Specific PLGA-DY-635 Nanoparticles Loaded with the Protein Kinase C Inhibitor Bisindolylmaleimide I

Pharmaceutics ◽

10.3390/pharmaceutics12111110 ◽

2020 ◽

Vol 12 (11) ◽

pp. 1110

Author(s):

Blerina Shkodra ◽

Adrian T. Press ◽

Antje Vollrath ◽

Ivo Nischang ◽

Stephanie Schubert ◽

...

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Targeted Delivery ◽

Near Infrared ◽

Water Solubility ◽

Analytical Ultracentrifugation ◽

Therapeutic Potential ◽

Kinase C ◽

Protein Kinase C Inhibitor

Bisindolylmaleimide I (BIM-I) is a competitive pan protein kinase C inhibitor with anti-inflammatory and anti-metastatic properties, suggested to treat inflammatory diseases and various cancer entities. However, despite its therapeutic potential, BIM-I has two major drawbacks, i.e., it has a poor water solubility, and it binds the human ether-à-go-go-related gene (hERG) ion channels, potentially causing deadly arrhythmias. In this case, a targeted delivery of BIM-I is imperative to minimize peripheral side effects. To circumvent these drawbacks BIM-I was encapsulated into nanoparticles prepared from poly(lactic-co-glycolic acid) (PLGA) functionalized by the near-infrared dye DY-635. DY-635 served as an active targeting moiety since it selectively binds the OATP1B1 and OATP1B3 transporters that are highly expressed in liver and cancer cells. PLGA-DY-635 (BIM-I) nanoparticles were produced by nanoprecipitation and characterized using dynamic light scattering, analytical ultracentrifugation, and cryogenic transmission electron microscopy. Particle sizes were found to be in the range of 20 to 70 nm, while a difference in sizes between the drug-loaded and unloaded particles was observed by all analytical techniques. In vitro studies demonstrated that PLGA-DY-635 (BIM-I) NPs prevent the PKC activation efficiently, proving the efficacy of the inhibitor after its encapsulation, and suggesting that BIM-I is released from the PLGA-NPs. Ultimately, our results present a feasible formulation strategy that improved the cytotoxicity profile of BIM-I and showed a high cellular uptake in the liver as demonstrated in vivo by intravital microscopy investigations.

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Isolation and characterization of protein kinase C from Y-1 adrenal cell cytoskeleton.

The Journal of Cell Biology ◽

10.1083/jcb.108.2.553 ◽

1989 ◽

Vol 108 (2) ◽

pp. 553-567 ◽

Cited By ~ 39

Author(s):

V Papadopoulos ◽

P F Hall

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Light Chain ◽

Myosin Light Chain ◽

Phorbol Esters ◽

Intact Cells ◽

Adrenal Cells ◽

Isolation And Characterization ◽

Kinase C

The cytoskeletons of Y-1 mouse adrenal tumor cells contain a calcium and phospholipid-dependent protein kinase (protein kinase C) that is bound sufficiently tight to resist extraction by 0.5% Triton but not by 1.0% Triton. The enzyme has been purified to near homogeneity from cytoskeleton and cytosol. It shows features typical of this type of kinase, namely a requirement for Ca2+ and phospholipid, stimulation by tumor promoters but not by nontumor-promoting phorbol esters, and inhibition by trifluoperazine. The enzyme shows specificity for four substrates found in the cytoskeleton, namely 80, 33, 20, and 18 kD. The first three substrates are phosphorylated by the enzyme; the fourth is dephosphorylated and is therefore affected by the kinase indirectly. The 80-kD protein is the kinase enzyme itself which is autophosphorylated in vitro and in the cytoskeleton. The 20-kD protein is myosin light chain. The 33- and 18-kD proteins are unidentified. The same substrates were phosphorylated when Y-1 cells were permeabilized with digitonin and incubated with [gamma-32P]ATP and phorbol-12-myristate-13-acetate. Partly purified protein kinase C changes the extent of phosphorylation of the same substrates when added to cytoskeletons previously extracted to remove endogenous protein kinase C. Addition of Ca2+, phosphatidylserine, and phorbol-12-myristate-13-acetate to cytoskeletons, and addition of these three agents plus protein kinase C to extracted cytoskeletons, causes these structures to undergo a rapid and extensive rounding. A similar change is induced in intact cells by addition of phorbol ester. It is concluded that protein kinase C is capable of changing the shape of adrenal cells by an action that involves autophosphorylation and phosphorylation of myosin light chain. This response may in turn be related to the steroidogenic responses to ACTH and cyclic AMP.

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Protein kinase C-linked inactivation of the interleukin-1 receptor in a human transformed B-cell line

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research ◽

10.1016/0167-4889(90)90073-m ◽

1990 ◽

Vol 1052 (1) ◽

pp. 173-178 ◽

Cited By ~ 7

Author(s):

Richard Horuk ◽

Janet L. Gross

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

B Cell ◽

Cell Line ◽

Interleukin 1 ◽

Kinase C

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Characterization of the Rac-GAP (Rac-GTPase-activating protein) activity of β2-chimaerin, a ‘non-protein kinase C’ phorbol ester receptor

Biochemical Journal ◽

10.1042/bj20030727 ◽

2003 ◽

Vol 375 (2) ◽

pp. 313-321 ◽

Cited By ~ 71

Author(s):

Maria Jose CALOCA ◽

HongBin WANG ◽

Marcelo G. KAZANIETZ

Keyword(s):

Protein Kinase C ◽

Protein Kinase ◽

Phorbol Ester ◽

Phorbol Esters ◽

Gtpase Activating Protein ◽

Protein Activity ◽

Rac Gtpase ◽

Kinase C ◽

Phorbol Ester Receptor

The regulation and function of β2-chimaerin, a novel receptor for the phorbol ester tumour promoters and the second messenger DAG (diacylglycerol), is largely unknown. As with PKC (protein kinase C) isoenzymes, phorbol esters bind to β2-chimaerin with high affinity and promote its subcellular distribution. β2-Chimaerin has GAP (GTPase-activating protein) activity for the small GTP-binding protein Rac1, but for not Cdc42 or RhoA. We show that acidic phospholipids enhanced its catalytic activity markedly in vitro, but the phorbol ester PMA had no effect. β2-Chimaerin and other chimaerin isoforms decreased cellular levels of Rac-GTP markedly in COS-1 cells and impaired GTP loading on to Rac upon EGF (epidermal growth factor) receptor stimulation. Deletional and mutagenesis analysis determined that the β2-chimaerin GAP domain is essential for this effect. Interestingly, PMA has a dual effect on Rac-GTP levels in COS-1 cells. PMA increased Rac-GTP levels in the absence of a PKC inhibitor, whereas under conditions in which PKC activity is inhibited, PMA markedly decreased Rac-GTP levels and potentiated the effect of β2-chimaerin. Chimaerin isoforms co-localize at the plasma membrane with active Rac, and these results were substantiated by co-immunoprecipitation assays. In summary, the novel phorbol ester receptor β2-chimaerin regulates the activity of the Rac GTPase through its GAP domain, leading to Rac inactivation. These results strongly emphasize the high complexity of DAG signalling due to the activation of PKC-independent pathways, and cast doubts regarding the selectivity of phorbol esters and DAG analogues as selective PKC activators.

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