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PMID: 15899855 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Keap1 regulates the oxidation-sensitive shuttling of Nrf2 into and out of the nucleus via a Crm1-dependent nuclear export mechanism.

Molecular and cellular biology ·Vol. 25 ·No. 11 ·2005-06-00 ·Pages 4501-13

Velichkova M, Hasson T

Abstract

Keap1 is a negative regulator of Nrf2, a transcription factor essential for antioxidant response element (ARE)-mediated gene expression. We find that Keap1 sequesters Nrf2 in the cytoplasm, not by docking it to the actin cytoskeleton but instead through an active Crm1/exportin-dependent nuclear export mechanism. Deletion and mutagenesis studies identified a nuclear export signal (NES) in the intervening region of Keap1 comprised of hydrophobic leucine and isoleucine residues in agreement with a traditional NES consensus sequence. Mutation of the hydrophobic amino acids resulted in nuclear accumulation of both Keap1 and Nrf2, as did treatment with the drug leptomycin B, which inactivates Crm1/exportin. ARE genes were partially activated under these conditions, suggesting that additional oxidation-sensitive elements are required for full activation of the antioxidant response. Based on these data, we propose a new model for regulation of Nrf2 by Keap1. Under normal conditions, Keap1 and Nrf2 are complexed in the cytoplasm where they are targeted for degradation. Oxidative stress inactivates Keap1's NES, allowing entry of both Keap1 and Nrf2 into the nucleus and transcriptional transactivation of ARE genes.

MeSH Terms
Active Transport, Cell Nucleus Adaptor Proteins, Signal Transducing/analysis,genetics,physiology Amino Acid Sequence Animals Antioxidants/metabolism Cell Nucleus/chemistry,metabolism Cytoplasm/chemistry,metabolism Cytoskeletal Proteins/analysis,genetics,physiology Cytoskeleton/metabolism DNA-Binding Proteins/analysis,metabolism Fatty Acids, Unsaturated/pharmacology Humans Karyopherins/metabolism,physiology Kelch-Like ECH-Associated Protein 1 Mice Molecular Sequence Data Mutation NF-E2-Related Factor 2 Nuclear Localization Signals/genetics,physiology Oxidation-Reduction Oxidative Stress/physiology Rats Receptors, Cytoplasmic and Nuclear/metabolism,physiology Response Elements Trans-Activators/analysis,metabolism
Chemicals
Adaptor Proteins, Signal Transducing Antioxidants Cytoskeletal Proteins DNA-Binding Proteins Fatty Acids, Unsaturated Karyopherins Keap1 protein, mouse Kelch-Like ECH-Associated Protein 1 NF-E2-Related Factor 2 NFE2L2 protein, human Nfe2l2 protein, mouse Nfe2l2 protein, rat Nuclear Localization Signals Receptors, Cytoplasmic and Nuclear Trans-Activators exportin 1 protein leptomycin B
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Velichkova Michaella
University of California at San Diego, Division of Biological Sciences, Section of Cell and Developmental Biology, 2129 Bonner Hall, MC 0368, 9500 Gilman Drive, La Jolla, CA 92093-0368, USA. velichko@biomail.ucsd.edu
Hasson Tama
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.11.4501-4513.2005
Email
velichko@biomail.ucsd.edu
Published
2005-06-00
Pages
4501-13
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1140621
Subset
IM
Grants
NEI NIH HHS · R01 EY012695 · United States
NEI NIH HHS · R01-EY12695 · United States
External Links
PubMed source DOI: 10.1128/MCB.25.11.4501-4513.2005 PMC Full Text View journal details
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