The NHB1 (N-terminal homology box 1) sequence in transcription factor Nrf1 is required to anchor it to the endoplasmic reticulum and also to enable its asparagine-glycosylation

Yiguo Zhang, John M. Lucocq, Masayuki Yamamoto, John D. Hayes

    Research output: Contribution to journalArticle

    58 Citations (Scopus)

    Abstract

    Nrf1 (nuclear factor-erythroid 2 p45 subunit-related factor 1) is negatively controlled by its NTD (N-terminal domain) that lies between amino acids 1 and 124. This domain contains a leucine-rich sequence, called NHB1 (N-terminal homology box 1; residues 11-30), which tethers Nrf1 to the ER (endoplasmic reticulum). Electrophoresis resolved Nrf1 into two major bands of approx. 95 and 120 kDa. The 120-kDa Nrf1 form represents a glycosylated protein that was present exclusively in the ER and was converted into a substantially smaller polypeptide upon digestion with either peptide:N-glycosidase F or endoglycosidase H. By contrast, the 95-kDa Nrf1 form did not appear to be glycosylated and was present primarily in the nucleus. NHB1 and its adjacent residues conform to the classic tripartite signal peptide sequence, comprising n-, h- and c-regions. The h-region (residues 11-22), but neither the n-region (residues 1-10) nor the c-region (residues 23-30), is required to direct Nrf1 to the ER. Targeting Nrf1 to the ER is necessary to generate the 120-kDa glycosylated protein. The n-region and c-region are required for correct membrane orientation of Nrf1, as deletion of residues 2-10 or 23-30 greatly increased its association with the ER and the extent to which it was glycosylated. The NHB1 does not contain a signal peptidase cleavage site, indicating that it serves as an ER anchor sequence. Wild-type Nrf1 is glycosylated through its Asn/Ser/Thr-rich domain, between amino acids 296 and 403, and this modification was not observed in an Nrf1(Delta299-400) mutant. Glycosylation of Nrf1 was not necessary to retain it in the ER.
    Original languageEnglish
    Pages (from-to)161-172
    Number of pages12
    JournalBiochemical Journal
    Volume408
    Issue number2
    DOIs
    Publication statusPublished - 2007

    Keywords

    • endoplasmic reticulum
    • glycosylation
    • non-alcoholic steatohepatitis (NASH)
    • nuclear factor-erythroid 2-p45 subunit-related factor (Nrf)
    • oxidative stress
    • signal anchor sequence
    • REGULATED INTRAMEMBRANE PROTEOLYSIS
    • ANTIOXIDANT RESPONSE ELEMENT
    • GENE-EXPRESSION
    • MEMBRANE-PROTEINS
    • INDUCIBLE EXPRESSION
    • EMBRYONIC LETHALITY
    • NEGATIVE REGULATION
    • SIGNAL SEQUENCES
    • ALPHA-COP
    • LIVER

    Cite this

    @article{71dd0c4d3f1440f6bc9f2e645f5e50a8,
    title = "The NHB1 (N-terminal homology box 1) sequence in transcription factor Nrf1 is required to anchor it to the endoplasmic reticulum and also to enable its asparagine-glycosylation",
    abstract = "Nrf1 (nuclear factor-erythroid 2 p45 subunit-related factor 1) is negatively controlled by its NTD (N-terminal domain) that lies between amino acids 1 and 124. This domain contains a leucine-rich sequence, called NHB1 (N-terminal homology box 1; residues 11-30), which tethers Nrf1 to the ER (endoplasmic reticulum). Electrophoresis resolved Nrf1 into two major bands of approx. 95 and 120 kDa. The 120-kDa Nrf1 form represents a glycosylated protein that was present exclusively in the ER and was converted into a substantially smaller polypeptide upon digestion with either peptide:N-glycosidase F or endoglycosidase H. By contrast, the 95-kDa Nrf1 form did not appear to be glycosylated and was present primarily in the nucleus. NHB1 and its adjacent residues conform to the classic tripartite signal peptide sequence, comprising n-, h- and c-regions. The h-region (residues 11-22), but neither the n-region (residues 1-10) nor the c-region (residues 23-30), is required to direct Nrf1 to the ER. Targeting Nrf1 to the ER is necessary to generate the 120-kDa glycosylated protein. The n-region and c-region are required for correct membrane orientation of Nrf1, as deletion of residues 2-10 or 23-30 greatly increased its association with the ER and the extent to which it was glycosylated. The NHB1 does not contain a signal peptidase cleavage site, indicating that it serves as an ER anchor sequence. Wild-type Nrf1 is glycosylated through its Asn/Ser/Thr-rich domain, between amino acids 296 and 403, and this modification was not observed in an Nrf1(Delta299-400) mutant. Glycosylation of Nrf1 was not necessary to retain it in the ER.",
    keywords = "endoplasmic reticulum, glycosylation, non-alcoholic steatohepatitis (NASH), nuclear factor-erythroid 2-p45 subunit-related factor (Nrf), oxidative stress, signal anchor sequence, REGULATED INTRAMEMBRANE PROTEOLYSIS, ANTIOXIDANT RESPONSE ELEMENT, GENE-EXPRESSION, MEMBRANE-PROTEINS, INDUCIBLE EXPRESSION, EMBRYONIC LETHALITY, NEGATIVE REGULATION, SIGNAL SEQUENCES, ALPHA-COP, LIVER",
    author = "Yiguo Zhang and Lucocq, {John M.} and Masayuki Yamamoto and Hayes, {John D.}",
    year = "2007",
    doi = "10.1042/BJ20070761",
    language = "English",
    volume = "408",
    pages = "161--172",
    journal = "Biochemical Journal",
    issn = "0264-6021",
    publisher = "Portland Press",
    number = "2",

    }

    The NHB1 (N-terminal homology box 1) sequence in transcription factor Nrf1 is required to anchor it to the endoplasmic reticulum and also to enable its asparagine-glycosylation. / Zhang, Yiguo; Lucocq, John M.; Yamamoto, Masayuki; Hayes, John D.

    In: Biochemical Journal, Vol. 408, No. 2, 2007, p. 161-172.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The NHB1 (N-terminal homology box 1) sequence in transcription factor Nrf1 is required to anchor it to the endoplasmic reticulum and also to enable its asparagine-glycosylation

    AU - Zhang, Yiguo

    AU - Lucocq, John M.

    AU - Yamamoto, Masayuki

    AU - Hayes, John D.

    PY - 2007

    Y1 - 2007

    N2 - Nrf1 (nuclear factor-erythroid 2 p45 subunit-related factor 1) is negatively controlled by its NTD (N-terminal domain) that lies between amino acids 1 and 124. This domain contains a leucine-rich sequence, called NHB1 (N-terminal homology box 1; residues 11-30), which tethers Nrf1 to the ER (endoplasmic reticulum). Electrophoresis resolved Nrf1 into two major bands of approx. 95 and 120 kDa. The 120-kDa Nrf1 form represents a glycosylated protein that was present exclusively in the ER and was converted into a substantially smaller polypeptide upon digestion with either peptide:N-glycosidase F or endoglycosidase H. By contrast, the 95-kDa Nrf1 form did not appear to be glycosylated and was present primarily in the nucleus. NHB1 and its adjacent residues conform to the classic tripartite signal peptide sequence, comprising n-, h- and c-regions. The h-region (residues 11-22), but neither the n-region (residues 1-10) nor the c-region (residues 23-30), is required to direct Nrf1 to the ER. Targeting Nrf1 to the ER is necessary to generate the 120-kDa glycosylated protein. The n-region and c-region are required for correct membrane orientation of Nrf1, as deletion of residues 2-10 or 23-30 greatly increased its association with the ER and the extent to which it was glycosylated. The NHB1 does not contain a signal peptidase cleavage site, indicating that it serves as an ER anchor sequence. Wild-type Nrf1 is glycosylated through its Asn/Ser/Thr-rich domain, between amino acids 296 and 403, and this modification was not observed in an Nrf1(Delta299-400) mutant. Glycosylation of Nrf1 was not necessary to retain it in the ER.

    AB - Nrf1 (nuclear factor-erythroid 2 p45 subunit-related factor 1) is negatively controlled by its NTD (N-terminal domain) that lies between amino acids 1 and 124. This domain contains a leucine-rich sequence, called NHB1 (N-terminal homology box 1; residues 11-30), which tethers Nrf1 to the ER (endoplasmic reticulum). Electrophoresis resolved Nrf1 into two major bands of approx. 95 and 120 kDa. The 120-kDa Nrf1 form represents a glycosylated protein that was present exclusively in the ER and was converted into a substantially smaller polypeptide upon digestion with either peptide:N-glycosidase F or endoglycosidase H. By contrast, the 95-kDa Nrf1 form did not appear to be glycosylated and was present primarily in the nucleus. NHB1 and its adjacent residues conform to the classic tripartite signal peptide sequence, comprising n-, h- and c-regions. The h-region (residues 11-22), but neither the n-region (residues 1-10) nor the c-region (residues 23-30), is required to direct Nrf1 to the ER. Targeting Nrf1 to the ER is necessary to generate the 120-kDa glycosylated protein. The n-region and c-region are required for correct membrane orientation of Nrf1, as deletion of residues 2-10 or 23-30 greatly increased its association with the ER and the extent to which it was glycosylated. The NHB1 does not contain a signal peptidase cleavage site, indicating that it serves as an ER anchor sequence. Wild-type Nrf1 is glycosylated through its Asn/Ser/Thr-rich domain, between amino acids 296 and 403, and this modification was not observed in an Nrf1(Delta299-400) mutant. Glycosylation of Nrf1 was not necessary to retain it in the ER.

    KW - endoplasmic reticulum

    KW - glycosylation

    KW - non-alcoholic steatohepatitis (NASH)

    KW - nuclear factor-erythroid 2-p45 subunit-related factor (Nrf)

    KW - oxidative stress

    KW - signal anchor sequence

    KW - REGULATED INTRAMEMBRANE PROTEOLYSIS

    KW - ANTIOXIDANT RESPONSE ELEMENT

    KW - GENE-EXPRESSION

    KW - MEMBRANE-PROTEINS

    KW - INDUCIBLE EXPRESSION

    KW - EMBRYONIC LETHALITY

    KW - NEGATIVE REGULATION

    KW - SIGNAL SEQUENCES

    KW - ALPHA-COP

    KW - LIVER

    U2 - 10.1042/BJ20070761

    DO - 10.1042/BJ20070761

    M3 - Article

    VL - 408

    SP - 161

    EP - 172

    JO - Biochemical Journal

    JF - Biochemical Journal

    SN - 0264-6021

    IS - 2

    ER -