Abstract
NUCLEAR factor κB" (NF-κB"), which was first detected by its binding to the κB" site in the immunoglobulin κ-gene enhancer1, is important for the regulated expression of the κ-gene2,3 and is partly responsible for the induction in appropriate cells of inter-leukin-2 (IL-2) 4, IL-2α receptor5, β-interferon6,7 and serum amyloid A protein8. NF-κ" is present as a nuclear DNA-binding protein in B lymphocytes and mature macrophages, but is found in the cytoplasm of many cells in a form unable to bind to DNA9. The cytoplasmic form is bound to an inhibitor protein, IκB, from which it can be released in vitro by deoxycholate and other agents10. Activation of cells by various agents, notably the phorbol esters that stimulate protein kinase C (PKC), leads to dissociation in vivo of the NF-κB/IκB complex and migration of NF-κB to the nucleus9. Therefore, it acts as a second messenger system, transducing activation signals from the cytoplasm to the nucleus. To elucidate the mechanism of signal transfer, we have used an in vitro system in which addition ofpurified protein kinases to a partially purified NF-κB/IκB complex leads to the activation of the DNA-binding activity of NF-κB. Using gel retardation assays we found that PKC, cyclic AMP-dependent protein kinase (PKA) and a haem-regulated eIF-2 kinase (HRI) could activate NF-κB in vitro, whereas casein kinase II was ineffective. To determine the target for the protein kinases we purified and characterized both NF-κB and IκB and found that IκB is phosphorylated and inactivated in the presence of PKC and HRI but not PKA.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Sen, R. & Baltimore, D. Cell 46, 705–716 (1986).
Lenardo, M. J., Pierce, J. W. & Baltimore, D. Science 236, 1573–1577 (1987).
Atchison, M. & Perry, R. P. Cell 48, 121–128 (1987).
Hoyos, B. et al. Science 244, 457–460 (1989).
Bohlnein, E. et al. Cell 53, 827–836 (1988).
Visvanathan, K. V. & Goodbourn, S. EMBO J. 8, 1129–1138 (1989).
Lenardo, M. J., Fan, C. M., Maniatis, T. & Baltimore, D. Cell 57, 287–294 (1989).
Edbrooke, M. R., Burt, D. W., Cheshire, J. K. & Woo, P. Molec. cell. Biol. 9, 1908–1916 (1989).
Sen, R. & Baltimore, D. Cell 47, 921–928 (1986).
Baeuerle, P. A. & Baltimore, D. Cell 53, 211–217 (1988).
Kikkawa, U. & Nishizuka, Y. A. Rev. Cell Biol. 2, 149–178 (1986).
Shirakawa, F. & Mizel, S. B. Molec. cell. Biol. 9, 2424–2430 (1989).
Woodgett, J. R. & Hunter, T., J. biol. Chem. 262, 4836–4843 (1987).
Huang, K-P., Nakabayashi, H. & Huang, F. L. Proc. natn. Acad. Sci. U.S.A. 83, 8535–8539 (1986).
Kawamoto, S. & Hidaka, H. Biochem. biophys. Res. Commun. 125, 258–264 (1984).
Knopf, J. L. et al. Cell 46, 491–502 (1986).
Moldave, K. A. Rev. Biochem. 54, 1109–1149 (1985).
Hathaway, G. M., Lundak, T. S., Tahara, S. M. & Traugh, J. A. Meth. Enzym. 60, 495–511 (1979).
Lenardo, M. J., Kuang, A., Gifford, A. & Baltimore, D. Proc. natn. Acad. Sci. U.S.A. 85, 8825–8829 (1988).
Kawakami, K., Scheidereit, C. & Roeder, R. G. Proc. natn. Acad. Sci. U.S.A. 85, 4700–4704 (1988).
Baeuerle, P. A. & Baltimore, D. Genes Dev. 3, 1689–1698 (1989).
Hager, D. A. & Burgess, R. R. Analyt. Biochem. 109, 76–78 (1980).
Baeuerle, P. A. & Baltimore, D. Science 242, 540–545 (1988).
Trachsel, H., Ranu, R. S. & London, I. M. Meth. Enzym. 60, 485–495 (1979).
Shirakawa, F. et al. Molec. cell. Biol. 9, 959–964 (1989).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ghosh, S., Baltimore, D. Activation in vitro of NF-κB" by phosphorylation of its inhibitor IκB". Nature 344, 678–682 (1990). https://doi.org/10.1038/344678a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/344678a0
This article is cited by
-
Understanding the functional role of membrane confinements in TNF-mediated signaling by multiscale simulations
Communications Biology (2022)
-
Metformin selectively dampens the acute inflammatory response through an AMPK-dependent mechanism
Scientific Reports (2021)
-
Deubiquitylating enzymes: potential target in autoimmune diseases
Inflammopharmacology (2021)
-
MicroRNA-mediated regulation of BM-MSCs differentiation into sweat gland-like cells: targeting NF-κB
Journal of Molecular Histology (2019)
-
Sanguinarine Attenuates Lipopolysaccharide-induced Inflammation and Apoptosis by Inhibiting the TLR4/NF-κB Pathway in H9c2 Cardiomyocytes
Current Medical Science (2018)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.