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PMID: 16872277 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Negative regulation of the Nrf1 transcription factor by its N-terminal domain is independent of Keap1: Nrf1, but not Nrf2, is targeted to the endoplasmic reticulum.

The Biochemical journal ·Vol. 399 ·No. 3 ·2006-11-01 ·Pages 373-85

Zhang Y, Crouch DH, Yamamoto M, Hayes JD

Abstract

Nrf1 (nuclear factor-erythroid 2 p45 subunit-related factor 1) and Nrf2 regulate ARE (antioxidant response element)-driven genes. At its N-terminal end, Nrf1 contains 155 additional amino acids that are absent from Nrf2. This 155-amino-acid polypeptide includes the N-terminal domain (NTD, amino acids 1-124) and a region (amino acids 125-155) that is part of acidic domain 1 (amino acids 125-295). Within acidic domain 1, residues 156-242 share 43% identity with the Neh2 (Nrf2-ECH homology 2) degron of Nrf2 that serves to destabilize this latter transcription factor through an interaction with Keap1 (Kelch-like ECH-associated protein 1). We have examined the function of the 155-amino-acid N-terminal polypeptide in Nrf1, along with its adjacent Neh2-like subdomain. Activation of ARE-driven genes by Nrf1 was negatively controlled by the NTD (N-terminal domain) through its ability to direct Nrf1 to the endoplasmic reticulum. Ectopic expression of wild-type Nrf1 and mutants lacking either the NTD or portions of its Neh2-like subdomain into wild-type and mutant mouse embryonic fibroblasts indicated that Keap1 controls neither the activity of Nrf1 nor its subcellular distribution. Immunocytochemistry showed that whereas Nrf1 gave primarily cytoplasmic staining that was co-incident with that of an endoplasmic-reticulum marker, Nrf2 gave primarily nuclear staining. Attachment of the NTD from Nrf1 to the N-terminus of Nrf2 produced a fusion protein that was redirected from the nucleus to the endoplasmic reticulum. Although this NTD-Nrf2 fusion protein exhibited less transactivation activity than wild-type Nrf2, it was nevertheless still negatively regulated by Keap1. Thus Nrf1 and Nrf2 are targeted to different subcellular compartments and are negatively regulated by distinct mechanisms.

MeSH Terms
Adaptor Proteins, Signal Transducing/deficiency,genetics,physiology Amino Acid Motifs Amino Acid Sequence Animals COS Cells Cell Nucleus/metabolism Cells, Cultured Chlorocebus aethiops Cytoskeletal Proteins/deficiency,genetics,physiology DNA, Complementary/genetics Endoplasmic Reticulum/metabolism Epithelial Cells Fibroblasts Genes, Reporter Humans Kelch-Like ECH-Associated Protein 1 Mice Mice, Knockout Molecular Sequence Data Mutagenesis, Site-Directed NF-E2-Related Factor 2/chemistry,genetics,metabolism Nuclear Proteins/metabolism Nuclear Respiratory Factor 1/chemistry,genetics,metabolism Protein Structure, Tertiary Protein Transport/physiology Rats Recombinant Fusion Proteins/metabolism Structure-Activity Relationship Transcriptional Activation
Chemicals
Adaptor Proteins, Signal Transducing Cytoskeletal Proteins DNA, Complementary Keap1 protein, mouse Kelch-Like ECH-Associated Protein 1 NF-E2-Related Factor 2 Nfe2l2 protein, mouse Nrf1 protein, mouse Nuclear Proteins Nuclear Respiratory Factor 1 Recombinant Fusion Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Zhang Yiguo
Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, Dundee DD1 9SY, Scotland, UK. y.z.zhang@dundee.ac.uk
Crouch Dorothy H
Yamamoto Masayuki
Hayes John D
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
1470-8728
DOI
10.1042/BJ20060725
Email
y.z.zhang@dundee.ac.uk
Published
2006-11-01
Pages
373-85
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1615900
Subset
IM
External Links
PubMed source DOI: 10.1042/BJ20060725 PMC Full Text View journal details
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