Effects of protease inhibitors on LPS-mediated activation of a mouse macrophage cell line (J774)

Abstract

Pretreatment (1 h) of a mouse macrophage-like cell line, J774, with the protease inhibitor, phenylalanine-chloromethyl ketone (PCK) or its structural analogue, tosylphenylalanine chloromethyl ketone (TPCK) was found to cause substantial inhibition of LPS-triggered activation of NF-κB. Pretreatment of cells with other types of protease inhibitors or their various structural analogues had no effect. PCK or TPCK appeared to exert its inhibitory effect by: (i) partially preventing LPS-triggered degradation of IκBα protein; (ii) preventing LPS-triggered nuclear translocation of NF-κB proteins (p50, RelA and Rel); and (iii) inhibiting the DNA-binding activities of NF-κB proteins. Pretreatment of cells with PCK or TPCK also resulted in the total or partial inhibition of LPS activatable (AP-1 or CREB) or constitutively-existing (Oct-1) transcription factors, but not of another constitutively-expressed transcription factor (SP-1). Pretreatment of J774 cells with PCK was found to substantially suppress LPS-induced expression of mRNAs specific for cytokine genes (TNFα, IL-1α and β, and IL-6), inducible nitric oxide synthase (iNOS) gene and IκBα gene, but not NF-κB1 p105 gene or β-actin gene. Furthermore, PCK pretreatment inhibited, in a dose-dependent manner, LPS-triggered production of nitric oxide production and tumoricidal activity.

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References

Adams DO Molecular interactions in macrophage activation. Immunol Today 1989; 10: 33-36.

Ohmori Y. , Hamilton TA Regulation of macrophage gene expression by T-cell-derived lymphokines. Pharmacol Ther 1994: 63: 235-264.

Muroi M. , Suzuki T. Role of protein kinase A in activation of NF-κB proteins of a mouse macrophage-like cell line, J774. Cell Signal 1993: 5: 289-298.

Muroi M. Muroi Y. , Yamamoto K. , Suzuki T. Influence of 3' half-site sequence of NF-κB motifs on the binding of lipopolysaccharide-activatable macrophage NF-κB proteins. J Biol Chem 1993: 268: 19534-19539.

Beg AA , Baldwin Jr AS The IκB proteins: multifunctional regulators of Rel/NF-κB transcription factors. Genes Dev 1993; 78: 2064-2070.

Baeuerle PA , Henkel T. Function and activation of NF-κB in the immune system. Annu Rev Immunol 1994; 12: 141-179.

Sen R. Baltimore D. Inducibility of κ immunoglobulin enhancer-binding protein NF-KB by a posttranslational mechanism. Cell 1986; 47: 921-928.

Ghosh S. , Baltimore D. Activation in vitro of NF-κB by phosphorylation of its inhibitor IκB . Nature 1990; 344: 678-682.

Shirakawa F. , Mizel SB In vitro activation and nuclear translocation of NF-κB catalyzed by cyclic AMP-dependent protein kinase and protein kinase C. Mol Cell Biol 1989; 9: 2424-2430.

10.

Shirakawa F. , Chedid M. , Suttles J. , Pollok BA , Mizel SB Interleukin 1 and cyclic AMP induce κ immunoglobulin light chain expression via activation of an NF-KB-like DNA-binding protein. Mol Cell Biol 1989; 9: 959-964.

11.

Meichle A. Schütze S. , Hensel G. , Brunsing D. , Krönke M. Protein kinase C-independent activation of nuclear factor KB by tumor necrosis factor. J Biol Chem 1990; 265: 8339-8343.

12.

Vinceti MP , Burrell TA , Taffet SM Regulation of NF-KB activity in murine macrophages: effect of bacterial lipopolysaccharide and phorbol ester. J Cell Physiol 1992; 150: 204-213.

13.

Finco TS , Baldwin Jr AS KB site-dependent induction of gene expression by diverse inducers of nuclear factor KB requires Raf-1. J Biol Chem 1993 ; 268: 17676-17679.

14.

Weinstein SL , June CH , DeFranco AL Lipopolysaccharide-induced protein tyrosine phosphorylation in human macrophages is mediated by CD14. J Immunol 1993; 151: 3829-3838.

15.

Beg AA , Baldwin Jr AS The IκB proteins: multifunctional regulators of Rel/NF-κB transcription factors. Genes Dev 1993; 78:2064-2070.

16.

Henkel T. , Machleid T. , Alklay I. , Krönke M. , Ben-Neriah Y. , Baeuerle PA Rapid proteolysis of IκB-α is necessary for activation of transcription factor NF-κB. Nature 1993; 365: 182-185.

17.

Rice NR , Ernst M. In vivo control of NF-KB activation by IκBα. EMBO J 1993; 12:4685-4695.

18.

Brown K. , Park S. , Kanno T. , Franzosa G. , Siebenlist U. Mutual regulation of the transcriptional activator NF-KB and its inhibitor, IκB-α. Proc Natl Acad Sci USA 1993 : 90: 2532-2536.

19.

Sun S-C. , Ganchi PA , Ballard DW , Greene WC NF-KB controls expression of inhibitor IκBα: evidence for an inducible autoregulatory pathway: Science 1993; 259: 1912-1915.

20.

Beg AA , Finco TS , Nantermet PV , Baldwin Jr AS Tumor necrosis factor and interleukin-1 lead to phosphorylation and loss of IκBα: a mechanism for NF-KB activation. Mol Cell Biol 1993; 13: 3301-3310.

21.

Cordle SR , Donald R. , Read MA , Hawiger J. Lipopolysaccharide induces phosphorylation of MAD3 and activation of Rel and related NF-KB proteins in human monocytic THP-1 cells. J Biol Chem 1993; 268: 11803-11810.

22.

Finco TS , Beg AA , Baldwin Jr AS Inducible phosphorylation of IκBα is not sufficient for its dissociation from NF-κB and is inhibited by protease inhibitors. Proc Natl Acad Sci USA 1994; 91: 11884-11888.

23.

Chiao PJ , Miyamoto S. , Verma IM Autoregulation of IκBα activity. Proc Natl Acad Sci USA 1994; 91: 28-32.

24.

Mellitis KH , Hay RT , Goodbourn S. Proteolytic degradation of MAD3 (IκBα) and enhanced processing of the NF-κB precursor p 105 are obligatory steps in the activation of NF-κB. Nucleic Acids Res 1993; 21: 5059-5066.

25.

Mackman N. Protease inhibitors block lipopolysaccharide induction of tissue factor gene expression in human monocytic cells by preventing activation of c-Rel/p65 heterodimers. J Biol Chem 1994; 269: 26363-26367.

26.

Palombella VJ , Rando OJ , Goldberg AL , Maniatis T. The ubiquitin-proteasome pathway is required for processing the NF-κB1 precursor protein and the activation of NF-κB . Cell 1994; 78: 773-785.

27.

Traenckner EB , Wilk S. , Baeuerle PA A proteasome inhibitor prevents activation of NF-κB and stabilizes a newly phosphorylated form of IKB-α that is still bound to NF-KB. EMBO J 1994; 13: 5433-5441.

28.

Ralph P. , Prichard J. , Cohen M. Reticulum cell sarcoma. An effector cell in antibody-dependent cell mediated immunity. J Immunol 1975; 114: 898-905.

29.

Fujihara M. , Muroi M. , Muroi Y. , Ito N. , Suzuki T. Mechanism of lipopolysaccharide-triggered junB activation in a mouse macrophage-like cell line (J774). J Biol Chem 1993; 268: 14898-14905.

30.

Muroi M. , Muroi Y. , Suzuki T. The binding of immobilized IgG2a to Fcγ2a receptor activates NF-κB via reactive oxygen intermediates and tumor necrosis factor-α. J Biol Chem 1994; 269: 30561-30568.

31.

Johnson WJ , Adams DO Assays detecting the antibody-dependent and independent binding and cytolysis of tumor cells by murine macrophages. Meth Enzymol 1986; 132: 555-568.

32.

Bressler P. , Brown K. , Timmer W. , Bours V. , Sibenlist U. , Fauci AS Mutational analysis of the p50 subunit of NF-κB and inhibition of NF-κB activity by trans-dominant p50 mutants . J Virol 1993; 67: 288-293.

33.

Rattner A. , Korner M. , Rosen H. , Baeuerle PA , Citri Y. Nuclear factor KB activates proenkephalin transcription in T lymphocytes. Mol Cell Biol 1991; 11: 1017-1022.

34.

Dignam JD , Lebowitz RM , Roeder RG Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 1983; 11: 1475-1489.

35.

Chomczynski P. , Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chl'oroform extraction. Anal Biochem 1987; 137: 156-159.

36.

Feinberg AP , Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132: 6-13.

37.

Khalili K. , Weinmann R. Shut-off of actin biosynthesis in adenovirus serotype-2-infected cells. J Biol Chem 1984; 258: 6701-6704.

38.

Miyamoto S. , Chiao PJ , Verma IM Enhanced IκBα degradation is responsible for constitutive NF-κB activity in murine B-cellline. Mol Cell Biol 1994; 14: 3276-3282.