Ubiquitination of Keap1, a BTB-Kelch substrate adaptor protein for Cul3, targets Keap1 for degradation by a proteasome-independent pathway

Donna Zhang, Shih Ching Lo, Zheng Sun, Geetha M. Habib, Michael W. Lieberman, Mark Hannink

Research output: Contribution to journalArticle

195 Citations (Scopus)

Abstract

Keap1 is a BTB-Kelch protein that functions as a substrate adaptor protein for a Cul3-dependent E3 ubiquitin ligase complex. Keap1 targets its substrate, the Nrf2 transcription factor, for ubiquitination and subsequent degradation by the 26 S proteasome. Inhibition of Keap1-dependent ubiquitination of Nrf2 increases steady-state levels of Nrf2 and enables activation of cytoprotective Nrf2-dependent genes. In this report, we demonstrate that Keap1 and three other BTB-Kelch proteins, including GAN1, ENC1, and Sarcosin, are ubiquitinated by a Cul3-dependent complex. Ubiquitination of Keap1 is markedly increased in cells exposed to quinone-induced oxidative stress, occurs in parallel with inhibition of Keap1-dependent ubiquitination of Nrf2, and results in decreased steady-state levels of Keap1, particularly in cells that are unable to synthesize glutathione. Degradation of Keap1 is independent of the 26 S proteasome, because inhibitors of the 26 S proteasome do not prevent loss of Keap1 following exposure of cells to quinone-induced oxidative stress. Our results suggest that a switch from substrate to substrate adaptor ubiquitination is a critical regulatory step that controls steady-state levels of both BTB-Kelch substrate adaptor proteins and their cognate substrates.

Original languageEnglish (US)
Pages (from-to)30091-30099
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number34
DOIs
StatePublished - Aug 26 2005
Externally publishedYes

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Ubiquitination
Proteasome Endopeptidase Complex
Degradation
Substrates
Proteins
Oxidative stress
Oxidative Stress
Proteasome Inhibitors
Ubiquitin-Protein Ligases
Glutathione
Transcription Factors
Genes
Chemical activation
Switches
benzoquinone

ASJC Scopus subject areas

  • Biochemistry

Cite this

Ubiquitination of Keap1, a BTB-Kelch substrate adaptor protein for Cul3, targets Keap1 for degradation by a proteasome-independent pathway. / Zhang, Donna; Lo, Shih Ching; Sun, Zheng; Habib, Geetha M.; Lieberman, Michael W.; Hannink, Mark.

In: Journal of Biological Chemistry, Vol. 280, No. 34, 26.08.2005, p. 30091-30099.

Research output: Contribution to journalArticle

Zhang, Donna ; Lo, Shih Ching ; Sun, Zheng ; Habib, Geetha M. ; Lieberman, Michael W. ; Hannink, Mark. / Ubiquitination of Keap1, a BTB-Kelch substrate adaptor protein for Cul3, targets Keap1 for degradation by a proteasome-independent pathway. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 34. pp. 30091-30099.
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