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Mol. Cell. Biol. 30 (13): 3275-3285

Copyright © 2010 by the American Society for Microbiology. All rights reserved.

A Noncanonical Mechanism of Nrf2 Activation by Autophagy Deficiency: Direct Interaction between Keap1 and p62{triangledown}

Alexandria Lau,1,{dagger} Xiao-Jun Wang,1,{dagger} Fei Zhao,1 Nicole F. Villeneuve,1 Tongde Wu,1 Tao Jiang,1 Zheng Sun,1 Eileen White,3,4, and Donna D. Zhang1,2*

Department of Pharmacology and Toxicology, University of Arizona, Tucson, Arizona 85721,1 Arizona Cancer Center, University of Arizona, Tucson, Arizona 85721,2 Cancer Institute of New Jersey, New Brunswick, New Jersey 08903,3 Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey 088544

Received for publication 3 March 2010. Accepted for publication 17 April 2010.

Abstract: In response to stress, cells can utilize several cellular processes, such as autophagy, which is a bulk-lysosomal degradation pathway, to mitigate damages and increase the chances of cell survival. Deregulation of autophagy causes upregulation of p62 and the formation of p62-containing aggregates, which are associated with neurodegenerative diseases and cancer. The Nrf2-Keap1 pathway functions as a critical regulator of the cell's defense mechanism against oxidative stress by controlling the expression of many cellular protective proteins. Under basal conditions, Nrf2 is ubiquitinated by the Keap1-Cul3-E3 ubiquitin ligase complex and targeted to the 26S proteasome for degradation. Upon induction, the activity of the E3 ubiquitin ligase is inhibited through the modification of cysteine residues in Keap1, resulting in the stabilization and activation of Nrf2. In this current study, we identified the direct interaction between p62 and Keap1 and the residues required for the interaction have been mapped to 349-DPSTGE-354 in p62 and three arginines in the Kelch domain of Keap1. Accumulation of endogenous p62 or ectopic expression of p62 sequesters Keap1 into aggregates, resulting in the inhibition of Keap1-mediated Nrf2 ubiquitination and its subsequent degradation by the proteasome. In contrast, overexpression of mutated p62, which loses its ability to interact with Keap1, had no effect on Nrf2 stability, demonstrating that p62-mediated Nrf2 upregulation is Keap1 dependent. These findings demonstrate that autophagy deficiency activates the Nrf2 pathway in a noncanonical cysteine-independent mechanism.

* Corresponding author. Mailing address: Department of Pharmacology and Toxicology, University of Arizona, 1703 East Mabel Street, Tucson, AZ 85721-0207. Phone: (520) 626-9918. Fax: (520) 626-2466. E-mail: dzhang{at}

{triangledown} Published ahead of print on 26 April 2010.

{dagger} Both authors contributed equally.

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