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

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

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.

scholarly journals 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.

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

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.

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

1991 ◽  
Vol 2 (4) ◽  
pp. 329-335 ◽  
Author(s):  
K Bomsztyk ◽  
J W Rooney ◽  
T Iwasaki ◽  
N A Rachie ◽  
S K Dower ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Cell Line ◽  
Phorbol Esters ◽  
Interleukin 1 ◽  
Interleukin 2 ◽  
Nf Kappa B ◽  
Kinase C ◽  
Protein Kinase C Inhibitor

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.

scholarly journals Role of intracellular calcium and protein kinase C in the endocytosis of transferrin and insulin by HL60 cells.

1986 ◽  
Vol 103 (3) ◽  
pp. 851-856 ◽  
Author(s):  
B Iacopetta ◽  
J L Carpentier ◽  
T Pozzan ◽  
D P Lew ◽  
P Gorden ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Myristate Acetate ◽  
Calcium Ionophore ◽  
Receptor Internalization ◽  
Minor Role ◽  
Myristate Acetate ◽  
Electron Microscope Autoradiography ◽  
Kinase C

The role of the cytosolic free calcium concentration ([Ca2+]i) and of protein kinase C on the internalization of transferrin and insulin in the human promyelocytic cell line HL60 was investigated. [Ca2+]i was selectively monitored and manipulated by the use of the fluorescent Ca2+ indicator and buffer quin2, while receptor-ligand internalization was studied directly by quantitative electron microscope autoradiography. Decreasing the [Ca2+]i up to 10-fold below resting level had no effect on the internalization of transferrin or insulin. Similarly, a 10-fold elevation of the [Ca2+]i using the calcium ionophore ionomycin caused little or no change in the endocytosis of the two ligands. In contrast, activation of protein kinase C by phorbol myristate acetate markedly stimulated the internalization of both occupied and unoccupied transferrin receptors, even in cells with very low [Ca2+]i. The insulin receptor was found to behave differently in response to phorbol myristate acetate, however, in that only the occupied receptors were stimulated to internalize. We conclude that the [Ca2+]i plays only a minor role in regulating receptor-mediated endocytosis, whereas protein kinase C can selectively modulate receptor internalization depending on receptor type and occupancy.

Regulation of paracellular permeability in Caco-2 cell monolayers by protein kinase C

1993 ◽  
Vol 265 (5) ◽  
pp. G955-G962 ◽  
Author(s):  
W. F. Stenson ◽  
R. A. Easom ◽  
T. E. Riehl ◽  
J. Turk
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Myristate Acetate ◽  
Paracellular Permeability ◽  
Colonic Adenocarcinoma ◽  
Myristate Acetate ◽  
Cell Monolayers ◽  
Kinase C ◽  
Endogenous Substrate ◽  
Stimulation Of

Caco-2 cells are an enterocyte-like cell line derived from a human colonic adenocarcinoma. Paracellular permeability was assessed in monolayers of these cells by transmonolayer resistance and by the permeation of [3H]mannitol across the monolayer. Paracellular permeability was increased by the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (50 nM), carbachol (500 microM), and the combination of carbachol (50 microM) and monolein (100 microM), an inhibitor of diacylglycerol kinase, as manifested by a decrease in transmonolayer resistance and an increase in mannitol permeation. The effects of all of these stimuli on transmonolayer resistance were inhibited by staurosporine (3 nM), an inhibitor of PKC. The effects of carbachol plus monolein were also inhibited by atropine (0.1 microM), a muscarinic antagonist. Treatment of the monolayers with each of the stimuli was associated with translocation of PKC activity from cytosol to a membrane-associated state. Stimulation of Caco-2 cell monolayers with phorbol myristate acetate or with the combination of carbachol and monolein was also associated with phosphorylation of the MARCKS protein, an endogenous substrate of PKC. These data support the hypothesis that intestinal paracellular permeability is regulated by the activity of enterocyte PKC and demonstrate that the increase in paracellular permeability induced by binding of carbachol to the muscarinic receptor is mediated by activation of PKC.

Involvement of protein kinase C in macrophage activation by poly(I.C)

1994 ◽  
Vol 266 (1) ◽  
pp. C134-C142 ◽  
Author(s):  
F. R. Lake ◽  
E. C. Dempsey ◽  
J. D. Spahn ◽  
D. W. Riches
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Myristate Acetate ◽  
Mrna Levels ◽  
Myristate Acetate ◽  
Western Blots ◽  
Level Of Involvement ◽  
Polycytidylic Acid ◽  
Kinase C ◽  
Polyinosinic Polycytidylic Acid

The expression of cytocidal activity is initiated by the interaction of macrophages with priming [e.g., interferon (IFN)] and triggering stimuli (polyinosinic-polycytidylic acid). We have shown that the triggering step can be initiated in a Ca(2+)-dependent fashion and hypothesized that protein kinase C (PKC) may couple the Ca2+ signal to the expression of a gene product, Bf, that accompanies the expression of macrophage cytocidal activity. Exposure of IFN-primed macrophages to polyinosinic-polycytidylic acid in the presence of the PKC inhibitors H-7 or sphingosine or after downregulation of PKC with phorbol myristate acetate markedly inhibited Bf synthesis. Western blots of macrophage lysates revealed the presence of the alpha-, delta-, and zeta-isozymes of PKC, and all were found to be downregulated by phorbol myristate acetate. Inhibition of PKC also prevented the increase in IFN-beta mRNA levels and partially blocked the response to IFN-beta. These data suggest that the alpha-, delta-, and zeta-isozymes of PKC are involved in signaling leading to Bf expression and that the level of involvement is restricted to the induction and response to IFN-beta.

Tetanus toxin attenuates the ability of phorbol myristate acetate to mobilize cytosolic protein kinase C in NG-108 cells

Toxicon ◽  
1990 ◽  
Vol 28 (1) ◽  
pp. 13-19 ◽  
Author(s):  
Robert V. Considine ◽  
John K. Bielicki ◽  
Lance L. Simpson ◽  
Joseph R. Sherwin
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Myristate Acetate ◽  
Tetanus Toxin ◽  
Myristate Acetate ◽  
Cytosolic Protein ◽  
Kinase C

Contrasting effects of inflammatory stimuli on neutrophil and monocyte adherence to endothelial cells

1989 ◽  
Vol 67 (2) ◽  
pp. 556-562 ◽  
Author(s):  
D. W. Kamp ◽  
K. D. Bauer ◽  
A. Knap ◽  
M. M. Dunn
Keyword(s):  
Hydrogen Peroxide ◽  
Endothelial Cells ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Myristate Acetate ◽  
Superoxide Anion ◽  
Leukocyte Adherence ◽  
Myristate Acetate ◽  
Kinase C ◽  
Monocyte Adherence

Leukocyte adherence to endothelial cells (EC) is an important early event in inflammatory responses, which are often characterized by a predominance of either neutrophils (PMN) or monocytes. However, there is little information concerning the molecular events important in leukocyte adherence to EC. Intracellular activation of protein kinase C and the calcium-second messenger system leads to the stimulation of a number of important functions in PMN and monocytes. We compared the effects of members of these pathways on human PMN and monocyte adherence to cultured bovine aortic EC. We observed that phorbol myristate acetate, phorbol, 12,13-dibutyrate, L-alpha-1-oleoyl-2-acetoyl-sn-3-glycerol, and ionomycin each induced significant dose-dependent increases in PMN adherence to EC monolayers. In contrast, similar concentrations of each of these agents induced significant decreases in EC adherence of monocytes enriched by countercurrent centrifugal elutriation. Separate experiments determined that the differences in PMN and monocyte adherence to EC were not related to differences in oxidant production because 1) phorbol myristate acetate and L-alpha-1-oleoyl-2-acetoyl-sn-3-glycerol caused similar marked increases in both PMN and monocyte superoxide anion and hydrogen peroxide production and 2) ionomycin, which had opposing effects on PMN and monocyte adherence, had no effect on PMN and monocyte superoxide anion or hydrogen peroxide release. We conclude that activators of protein kinase C and the Ca-second messenger pathway have opposite effects on PMN and monocyte adherence to EC and that these effects are mediated by O2 radical-independent mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS)

Sign in / Sign up

Export Citation Format

Share Document