Tumor necrosis factor activation of the sphingomyelin pathway signals nuclear factor κB translocation in intact HL-60 cells Journal Article
| Authors: | Yang, Z.; Costanzo, M.; Golde, D. W.; Kolesnick, R. N. |
| Article Title: | Tumor necrosis factor activation of the sphingomyelin pathway signals nuclear factor κB translocation in intact HL-60 cells |
| Abstract: | Recent investigations suggest that tumor necrosis factor (TNF)-a may utilize the sphingomyelin pathway for signal transduction. Signaling in this system involves hydrolysis of sphingomyelin to ceramide by action of a neutral sphingomyelinase and stimulation of a ceramide-activated protein kinase (Dressler, K. A., Mathias, S., and Kolesnick, R. N. (1992) Science 255, 1715-1718). To clarify the role of this pathway in TNF action, the present studies assessed the effect of the sphingomyelin pathway on activation of nuclear factor κB (NF-κB), an event considered integral to the transfer of the TNF message to the cell nucleus. As shown previously, TNF (1 nM) induced a marked increase in nuclear NF-κB binding in human leukemia (HL-60) cells within 5 min, and elevated binding was detected for as long as 1 h. Addition of a maximally effective concentration of sphingomyelinase, 0.1 units·ml-1, induced a 50% reduction in sphingomyelin content by 5 min from a basal level of 560 pmol·106 cells-1 and a quantitative increase in ceramide levels from 89 pmol·106 cells-1. Sphingomyelinase 0.1 units·ml-1 also induced an increase in nuclear NF-κB binding within 5 min, an effect measurable for as long as 1 h. As little as 1 × 10-5 units·ml-1 sphingomyelinase was effective and a maximal effect occurred with 1 × 10-3 units·ml-1. A cell-permeable ceramide analog, C8-ceramide, which mimics biologic effects of TNF-α, also enhanced nuclear NF-κB activation within minutes. In contrast, addition of a phospholipase C or a synthetic diacylglycerol (DG) analog, 1,2-dioctanoylglycerol, failed to enhance nuclear NF-κB binding despite large increases in cellular DG content. Further, TNF-α induced elevation in ceramide content by 2 min to 185% of control but did not affect DG levels. These studies provide evidence that stimulation of the sphingomyelin pathway leads to NF-κB activation in HL-60 cells. |
| Keywords: | signal transduction; human cell; protein binding; tumor cells, cultured; leukemia, promyelocytic, acute; molecular sequence data; tumor necrosis factor alpha; kinetics; nf-kappa b; base sequence; cell nucleus; receptor; cell stimulation; arachidonic acid; protein kinase; tumor necrosis factor; ceramide; ceramides; diglycerides; sphingomyelin phosphodiesterase; oligodeoxyribonucleotides; sphingomyelin; phospholipid metabolism; nuclear factor; enhancer elements (genetics); sphingomyelins; phospholipase c; human; priority journal; article; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; dioctanoin; phospholipases a |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 268 |
| Issue: | 27 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1993-09-25 |
| Start Page: | 20520 |
| End Page: | 20523 |
| Language: | English |
| PUBMED: | 8376408 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 1 March 2019 -- Source: Scopus |
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