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PMID: 9892650 Published · ppublish English Journal Article

Involvement of regulatory and catalytic subunits of phosphoinositide 3-kinase in NF-kappaB activation.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 2 ·1999-01-19 ·Pages 429-34

Béraud C, Henzel WJ, Baeuerle PA

Abstract

Hypoxia, reoxygenation, and the tyrosine phosphatase inhibitor pervanadate activate the transcription factor NF-kappaB, involving phosphorylation of its inhibitor IkappaB-alpha on tyrosine 42. This modification does not lead to degradation of IkappaB by the proteasome/ubiquitin pathway, as is seen on stimulation of cells with proinflammatory cytokines. It is currently unknown how tyrosine-phosphorylated IkappaB is removed from NF-kappaB. Here we show that p85alpha, the regulatory subunit of PI3-kinase, specifically associates through its Src homology 2 domains with tyrosine-phosphorylated IkappaB-alpha in vitro and in vivo after stimulation of T cells with pervanadate. This association could provide a mechanism by which newly tyrosine-phosphorylated IkappaB is sequestered from NF-kappaB. Another mechanism by which PI3-kinase contributed to NF-kappaB activation in response to pervanadate appeared to involve its catalytic p110 subunit. This was evident from the inhibition of pervanadate-induced NF-kappaB activation and reporter gene induction by treatment of cells with nanomolar amounts of the PI3-kinase inhibitor wortmannin. The compound had virtually no effect on tumor necrosis factor- and interleukin-1-induced NF-kappaB activities. Wortmannin did not inhibit tyrosine phosphorylation of IkappaB-alpha or alter the stability of the PI3-kinase complex but inhibited Akt kinase activation in response to pervanadate. Our data suggest that both the regulatory and the catalytic subunit of PI3-kinase play a role in NF-kappaB activation by the tyrosine phosphorylation-dependent pathway.

MeSH Terms
Androstadienes/pharmacology DNA-Binding Proteins/metabolism Enzyme Inhibitors/pharmacology Humans I-kappa B Proteins Jurkat Cells NF-kappa B/metabolism Peptides/metabolism Phosphatidylinositol 3-Kinases/metabolism Phosphorylation Phosphotyrosine/metabolism Transcriptional Activation/genetics Tumor Necrosis Factor-alpha/pharmacology Vanadates/pharmacology Wortmannin src Homology Domains/genetics
Chemicals
Androstadienes DNA-Binding Proteins Enzyme Inhibitors I-kappa B Proteins NF-kappa B Peptides Tumor Necrosis Factor-alpha pervanadate Phosphotyrosine Vanadates Phosphatidylinositol 3-Kinases Wortmannin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Béraud C
Tularik, Two Corporate Drive, South San Francisco, CA 94080, USA. beraud@cytokinetics.com
Henzel W J
Baeuerle P A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.2.429
Email
beraud@cytokinetics.com
Published
1999-01-19
Pages
429-34
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC15153
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.96.2.429 PMC Full Text View journal details
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