Mapping of the Inducible IκB Phosphorylation Sites That Signal Its Ubiquitination and Degradation†
Open accessAbstract
Extracellular stimuli that activate the transcription factor NF-kappaB cause rapid phosphorylation of the IkappaBalpha inhibitor, which retains NF-kappaB in the cytoplasm of nonstimulated cells. Phosphorylation of IkappaBalpha is followed by its rapid degradation, the inhibition of which prevents NF-kappaB activation. To determine the relationship between these events, we mapped the inducible phosphorylation sites of IkappaBalpha. We found that two residues, serines 32 and 36, were phosphorylated in response to either tumor necrosis factor, interleukin-1, or phorbol ester. Substitution of either serine blocks or slows down induction of IkappaBalpha degradation. Substitutions of the homologous sites in IkappaBbeta, serines 19 and 23, also prevent inducible IkappaBbeta degradation. We suggest that activation of a single IkappaB kinas e or closely related IkappaB kinases is the first cr itical step in NF-kappaB activation. Once phosphorylated, IkappaB is ubiquitinated. Unlike wild-type IkappaBalpha, the phosphorylation-defective mutants do not undergo inducible polyubiquitination. As substitution of a conserved lysine residue slows down the ubiquitination and degradation of IkappaBalpha without affecting its phosphorylation, polyubiquitination is required for inducible IkappaB degradation.
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- Authors
- Didonato, J., Mercurio, F., Rosette, C., Wu-Li, J., Suyang, H., Ghosh, S., & Karin, M.
- Affiliations
- University of California San Diego (US)
- Howard Hughes Medical Institute (US)
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 1996-04
- Volume
- 16
- Issue
- 4
- Pages
- 1295-1304
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC231113
- OpenAlex
- W2150033965
- Cited by
- 707
- References
- 51
- Citations by year
- 2026: 1 · 2025: 5 · 2024: 4 · 2023: 2 · 2022: 2 · 2021: 11 · 2020: 12 · 2019: 9 · 2018: 2 · 2017: 9 · 2016: 9 · 2015: 7 · 2014: 18 · 2013: 12 · 2012: 13
- FWCI
- 29.94
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Molecular and Cellular Biology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5p01ca050528-07)
- National Heart, Lung, and Blood Institute (HL 35018, P01HL035018)
- National Cancer Institute (CA50528)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
- Keywords
- IκBα, IκBβ, NF-κB activation, phosphorylation sites, ubiquitination, protein degradation, IκB kinase, TNF-α, interleukin-1, phorbol ester, transcriptional repression
- MeSH terms
- NF-KappaB Inhibitor alpha, Animals, DNA-Binding Proteins, HeLa Cells, Humans, Interleukin-1, Lysine, Mutation, Peptide Mapping, Phosphorylation, Precipitin Tests, Protease Inhibitors, Serine, Tumor Necrosis Factor-alpha, Ubiquitins, NF-kappa B, 3T3 Cells, Protein Serine-Threonine Kinases, I-kappa B Proteins, Mice, I-kappa B Kinase
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