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

Yiguo Zhang, Dorothy H. Crouch, Masayuki Yamamoto, John D. Hayes

    Research output: Contribution to journalArticle

    83 Citations (Scopus)

    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.
    Original languageEnglish
    Pages (from-to)373-385
    Number of pages13
    JournalBiochemical Journal
    Volume399
    Issue number3
    DOIs
    Publication statusPublished - Nov 2006

    Fingerprint

    Endoplasmic Reticulum
    Transcription Factors
    Amino Acids
    Antioxidant Response Elements
    Proteins
    Fusion reactions
    Genes
    Staining and Labeling
    Peptides
    Fibroblasts
    Transcriptional Activation
    Kelch-Like ECH-Associated Protein 1
    Chemical activation
    Immunohistochemistry

    Keywords

    • Antioxidant response element
    • Endoplasmic reticulum
    • Kelch-like ECH-associated protein 1 (Keap1)
    • Nuclear factor-erythroid 2 p45 subunit-related factor 1 (Nrf1)
    • Nuclear factor-erythroid 2 p45 subunit-related factor 2 (Nrf2)
    • Oxidative stress

    Cite this

    @article{917ce3bfe0e945b6b5c9e2f035679bb2,
    title = "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",
    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.",
    keywords = "Antioxidant response element, Endoplasmic reticulum, Kelch-like ECH-associated protein 1 (Keap1), Nuclear factor-erythroid 2 p45 subunit-related factor 1 (Nrf1), Nuclear factor-erythroid 2 p45 subunit-related factor 2 (Nrf2), Oxidative stress",
    author = "Yiguo Zhang and Crouch, {Dorothy H.} and Masayuki Yamamoto and Hayes, {John D.}",
    year = "2006",
    month = "11",
    doi = "10.1042/BJ20060725",
    language = "English",
    volume = "399",
    pages = "373--385",
    journal = "Biochemical Journal",
    issn = "0264-6021",
    publisher = "Portland Press",
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    }

    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. / Zhang, Yiguo; Crouch, Dorothy H.; Yamamoto, Masayuki; Hayes, John D.

    In: Biochemical Journal, Vol. 399, No. 3, 11.2006, p. 373-385.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Negative regulation of the Nrf1 transcription factor by its N-terminal domain is independent of Keap1

    T2 - Nrf1, but not Nrf2, is targeted to the endoplasmic reticulum

    AU - Zhang, Yiguo

    AU - Crouch, Dorothy H.

    AU - Yamamoto, Masayuki

    AU - Hayes, John D.

    PY - 2006/11

    Y1 - 2006/11

    N2 - 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.

    AB - 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.

    KW - Antioxidant response element

    KW - Endoplasmic reticulum

    KW - Kelch-like ECH-associated protein 1 (Keap1)

    KW - Nuclear factor-erythroid 2 p45 subunit-related factor 1 (Nrf1)

    KW - Nuclear factor-erythroid 2 p45 subunit-related factor 2 (Nrf2)

    KW - Oxidative stress

    U2 - 10.1042/BJ20060725

    DO - 10.1042/BJ20060725

    M3 - Article

    VL - 399

    SP - 373

    EP - 385

    JO - Biochemical Journal

    JF - Biochemical Journal

    SN - 0264-6021

    IS - 3

    ER -