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PMID: 9418849 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Novel IkappaB alpha proteolytic pathway in WEHI231 immature B cells.

Molecular and cellular biology ·Vol. 18 ·No. 1 ·1998-01-00 ·Pages 19-29

Miyamoto S, Seufzer BJ, Shumway SD

Abstract

The Rel/NF-kappaB family of transcription factors is sequestered in the cytoplasm of most mammalian cells by inhibitor proteins belonging to the IkappaB family. Degradation of IkappaB by a phosphorylation-dependent ubiquitin-proteasome (inducible) pathway is believed to allow nuclear transport of active Rel/NF-kappaB dimers. Rel/NF-kappaB (a p50-c-Rel dimer) is constitutively nuclear in murine B cells, such as WEHI231 cells. In these cells, p50, c-Rel, and IkappaB alpha are synthesized at high levels but only IkappaB alpha is rapidly degraded. We have examined the mechanism of IkappaB alpha degradation and its relation to constitutive p50-c-Rel activation. We demonstrate that all IkappaB alpha is found complexed with c-Rel protein in the cytoplasm. Additionally, rapid IkappaB alpha proteolysis is independent of but coexistent with the inducible pathway and can be inhibited by calcium chelators and some calpain inhibitors. Conditions that prevent degradation of IkappaB alpha also inhibit nuclear p50-c-Rel activity. Furthermore, the half-life of nuclear c-Rel is much shorter than that of the cytoplasmic form, underscoring the necessity for its continuous nuclear transport to maintain constitutive p50-c-Rel activity. We observed that IkappaB beta, another NF-kappaB inhibitor, is also complexed with c-Rel but slowly degraded by a proteasome-dependent process in WEHI231 cells. In addition, IkappaB beta is basally phosphorylated and cytoplasmic. We thus suggest that calcium-dependent IkappaB alpha proteolysis maintains nuclear transport of a p50-c-Rel heterodimer which in turn activates the synthesis of IkappaB alpha, p50, and c-Rel to sustain this dynamic process in WEHI231 B cells.

MeSH Terms
Animals B-Lymphocytes/metabolism Cell Line Mice NF-kappa B/antagonists & inhibitors,genetics Proto-Oncogene Proteins/genetics,metabolism Signal Transduction Transcription Factor RelB Transcription Factors
Chemicals
NF-kappa B Proto-Oncogene Proteins Relb protein, mouse Transcription Factors Transcription Factor RelB
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Miyamoto S
Department of Human Oncology, University of Wisconsin-Madison, 53792, USA. miyamoto@humonc.wisc.edu
Seufzer B J
Shumway S D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.1.19
Email
miyamoto@humonc.wisc.edu
Published
1998-01-00
Pages
19-29
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC121444
Subset
IM
Grants
NIGMS NIH HHS · T32GM07215 · United States
External Links
PubMed source DOI: 10.1128/MCB.18.1.19 PMC Full Text View journal details
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