DOI: 10.1126/science.284.5412.309. Positive and Negative Regulation of IκB Kinase Activity Through IKKβ Subunit Phosphorylation

Positive and Negative Regulation of IκB Kinase Activity Through IKKβ Subunit Phosphorylation

10.1126/science.284.5412.309

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

IκB [inhibitor of nuclear factor κB (NF-κB)] kinase (IKK) phosphorylates IκB inhibitory proteins, causing their degradation and activation of transcription factor NF-κB, a master activator of inflammatory responses. IKK is composed of three subunits—IKKα and IKKβ, which are highly similar protein kinases, and IKKγ, a regulatory subunit. In mammalian cells, phosphorylation of two sites at the activation loop of IKKβ was essential for activation of IKK by tumor necrosis factor and interleukin-1. Elimination of equivalent sites in IKKα, however, did not interfere with IKK activation. Thus, IKKβ, not IKKα, is the target for proinflammatory stimuli. Once activated, IKKβ autophosphorylated at a carboxyl-terminal serine cluster. Such phosphorylation decreased IKK activity and may prevent prolonged activation of the inflammatory response.

Bibliography

Delhase, M., Hayakawa, M., Chen, Y., & Karin, M. (1999). Positive and Negative Regulation of IÎºB Kinase Activity Through IKKÎ² Subunit Phosphorylation. Science, 284(5412), 309â313.

Authors 4

Mireille Delhase (first)
Makio Hayakawa (additional)
Yi Chen (additional)
Michael Karin (additional)

References 37 Referenced 732

10.1146/annurev.iy.12.040194.001041
Ghosh S., May M. J., Kopp E. B., ibid. 16, 225 (1998); / ibid. by Ghosh S. (1998)
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10.1146/annurev.immunol.14.1.649
10.1038/41493
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Malinin N. L., Boldin M. P., Kovalenko A. V., Wallach D., ibid. 385, 540 (1997). / ibid. by Malinin N. L. (1997)
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10.1016/S0092-8674(00)81842-5
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M. Karin and M. Delhase ibid. p. 9067.
Cells incubated with [ 32 P]orthophosphate (2 mCi/ml) for 5 hours were TNF-stimulated washed and harvested in lysis buffer. IKK was immunoprecipitated from precleared lysates using anti-IKKα as described (3). Immune complexes were washed with lysis buffer containing 2 M urea. Proteins were resolved by SDS–polyacrylamide gel electrophoresis (7.5%) and transferred to Immobilon-P membrane (Millipore) for autoradiography and immunoblot analysis. IKK assays were performed as described (3).
See supplementary Web Fig. 1A available at the Science Web site (www.sciencemag.org/feature/data/987145.shl).
10.1128/MCB.16.4.1295
10.1016/0076-6879(91)01013-R
Supplementary Web Fig. 1B in (15).
Single-letter abbreviations for the amino acid residues are as follows: A Ala; C Cys; D Asp; E Glu; F Phe; G Gly; H His; I Ile; K Lys; L Leu; M Met; N Asn; P Pro; Q Gln; R Arg; S Ser; T Thr; V Val; W Trp; and Y Tyr.
M. Delhase and G. Natoli unpublished results; D. Goeddel personal communication.
10.1126/science.284.5412.316
Z. Li W. M. Chu M. Karin in preparation.
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10.1056/NEJM199704103361506
Supplementary Web Fig. 2 in (15).
10.1002/j.1460-2075.1994.tb06361.x
; D. R. Alessi et al. ibid. p. 1610; M. Yan and D. J. Templeton J. Biol. Chem. 269 19067 (1994). (10.1016/S0021-9258(17)32275-5)
10.1096/fasebj.9.13.7557015
10.1016/S0960-9822(95)00277-6
10.1016/S0955-0674(97)80058-0
10.1002/j.1460-2075.1995.tb00037.x
We thank D. Rothwarf J. DiDonato and F. Mercurio for helpful discussion and advice J. W. Li G. Cadwell and J. Lewis for excellent technical assistance and B. Thompson for help with manuscript preparation. M.D. M.H. and Y.C. were supported by postdoctoral fellowships from the D. Collen Research Foundation (Belgium) Human Frontier Science Program and Tobacco-Related Disease Research Program respectively. Work was supported by grants from The National Institutes of Health. (R37 ES04151 and R01 AI43477) and the U.S. Department of Energy (DE-FG03-86ER60429). M.K. is a Frank and Else Schilling–American Cancer Society Research Professor.

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 5:40 a.m.)
Deposited	1 year, 7 months ago (Jan. 12, 2024, 10:48 p.m.)
Indexed	1 week, 3 days ago (Aug. 21, 2025, 1:41 p.m.)
Issued	26 years, 4 months ago (April 9, 1999)
Published	26 years, 4 months ago (April 9, 1999)
Published Print	26 years, 4 months ago (April 9, 1999)

Funders 0

None

BibTeX

@article{Delhase_1999, title={Positive and Negative Regulation of IκB Kinase Activity Through IKKβ Subunit Phosphorylation}, volume={284}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.284.5412.309}, DOI={10.1126/science.284.5412.309}, number={5412}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Delhase, Mireille and Hayakawa, Makio and Chen, Yi and Karin, Michael}, year={1999}, month=apr, pages={309–313} }

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  "abstract": "<jats:p>I\u03baB [inhibitor of nuclear factor \u03baB (NF-\u03baB)] kinase (IKK) phosphorylates I\u03baB inhibitory proteins, causing their degradation and activation of transcription factor NF-\u03baB, a master activator of inflammatory responses. IKK is composed of three subunits\u2014IKK\u03b1 and IKK\u03b2, which are highly similar protein kinases, and IKK\u03b3, a regulatory subunit. In mammalian cells, phosphorylation of two sites at the activation loop of IKK\u03b2 was essential for activation of IKK by tumor necrosis factor and interleukin-1. Elimination of equivalent sites in IKK\u03b1, however, did not interfere with IKK activation. Thus, IKK\u03b2, not IKK\u03b1, is the target for proinflammatory stimuli. Once activated, IKK\u03b2 autophosphorylated at a carboxyl-terminal serine cluster. Such phosphorylation decreased IKK activity and may prevent prolonged activation of the inflammatory response.</jats:p>",
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