Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter

Andreas Kolb, W H GUNZBURG, R ALBANG, G BREM, V ERFLE, B SALMONS

    Research output: Contribution to journalArticle

    21 Citations (Scopus)

    Abstract

    Expression of the whey acidic protein (WAP) gene is tightly regulated in a tissue and developmental stage specific manner, in that the WAP gene is exclusively expressed in the mammary gland during pregnancy and lactation. Using both deletion and competition analyses, evidence is provided for the existence of a negative regulatory element (NRE) in the WAP promoter located between -413 and -93 with respect to the WAP transcriptional initiation site. This NRE dramatically decreases transcription from linked heterologous promoter-reporter gene constructs. The activity of NRE requires WAP promoter sequences that are 230 bp apart since subfragments of the NRE fail to inhibit transcription of adjoining reporter genes. Nuclear extracts from different cell types, in which the WAP gene is not active, contain a protein or complex that specifically interacts with the entire NRE but not with subfragments of it. The contact points between this protein (NRE binding factor [NBF]) and the NRE element have been partially determined. Mutation of the implicated nucleotides severely reduces the ability of NBF to bind, and such mutated promoter fragments fail to alleviate transcriptional repression in competition experiments. This suggests that NBF binding to the NRE is at least in part responsible for the negative regulation of the WAP promoter. Since NBF is not detectable in the lactating mammary gland, where the WAP gene is expressed, we speculate that it may be a determinant of the expression spectrum of the WAP gene. (C) 1994 Wiley-Liss, Inc.

    Original languageEnglish
    Pages (from-to)245-261
    Number of pages17
    JournalJournal of Cellular Biochemistry
    Volume56
    Issue number2
    Publication statusPublished - Oct 1994

    Keywords

    • negative regulatory element
    • heterologous promoter
    • DNA binding factor
    • transcriptional repression
    • milk protein expression control
    • cell-specific enhancer
    • high-level expression
    • long terminal repeat
    • tumor virus
    • transgenic mice
    • gene-expression
    • nuclear factor
    • hormonal-regulation
    • upstream sequences

    Cite this

    Kolb, A., GUNZBURG, W. H., ALBANG, R., BREM, G., ERFLE, V., & SALMONS, B. (1994). Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter. Journal of Cellular Biochemistry, 56(2), 245-261.

    Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter. / Kolb, Andreas; GUNZBURG, W H ; ALBANG, R ; BREM, G ; ERFLE, V ; SALMONS, B .

    In: Journal of Cellular Biochemistry, Vol. 56, No. 2, 10.1994, p. 245-261.

    Research output: Contribution to journalArticle

    Kolb, A, GUNZBURG, WH, ALBANG, R, BREM, G, ERFLE, V & SALMONS, B 1994, 'Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter', Journal of Cellular Biochemistry, vol. 56, no. 2, pp. 245-261.
    Kolb A, GUNZBURG WH, ALBANG R, BREM G, ERFLE V, SALMONS B. Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter. Journal of Cellular Biochemistry. 1994 Oct;56(2):245-261.
    Kolb, Andreas ; GUNZBURG, W H ; ALBANG, R ; BREM, G ; ERFLE, V ; SALMONS, B . / Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter. In: Journal of Cellular Biochemistry. 1994 ; Vol. 56, No. 2. pp. 245-261.
    @article{80488bb5c5984a039cf7dca26bc0a7b3,
    title = "Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter",
    abstract = "Expression of the whey acidic protein (WAP) gene is tightly regulated in a tissue and developmental stage specific manner, in that the WAP gene is exclusively expressed in the mammary gland during pregnancy and lactation. Using both deletion and competition analyses, evidence is provided for the existence of a negative regulatory element (NRE) in the WAP promoter located between -413 and -93 with respect to the WAP transcriptional initiation site. This NRE dramatically decreases transcription from linked heterologous promoter-reporter gene constructs. The activity of NRE requires WAP promoter sequences that are 230 bp apart since subfragments of the NRE fail to inhibit transcription of adjoining reporter genes. Nuclear extracts from different cell types, in which the WAP gene is not active, contain a protein or complex that specifically interacts with the entire NRE but not with subfragments of it. The contact points between this protein (NRE binding factor [NBF]) and the NRE element have been partially determined. Mutation of the implicated nucleotides severely reduces the ability of NBF to bind, and such mutated promoter fragments fail to alleviate transcriptional repression in competition experiments. This suggests that NBF binding to the NRE is at least in part responsible for the negative regulation of the WAP promoter. Since NBF is not detectable in the lactating mammary gland, where the WAP gene is expressed, we speculate that it may be a determinant of the expression spectrum of the WAP gene. (C) 1994 Wiley-Liss, Inc.",
    keywords = "negative regulatory element, heterologous promoter, DNA binding factor, transcriptional repression, milk protein expression control, cell-specific enhancer, high-level expression, long terminal repeat, tumor virus, transgenic mice, gene-expression, nuclear factor, hormonal-regulation, upstream sequences",
    author = "Andreas Kolb and GUNZBURG, {W H} and R ALBANG and G BREM and V ERFLE and B SALMONS",
    year = "1994",
    month = "10",
    language = "English",
    volume = "56",
    pages = "245--261",
    journal = "Journal of Cellular Biochemistry",
    issn = "0730-2312",
    publisher = "Wiley-Liss Inc.",
    number = "2",

    }

    TY - JOUR

    T1 - Negative Regulatory Element in the Mammary Specific Whey Acidic Protein Promoter

    AU - Kolb, Andreas

    AU - GUNZBURG, W H

    AU - ALBANG, R

    AU - BREM, G

    AU - ERFLE, V

    AU - SALMONS, B

    PY - 1994/10

    Y1 - 1994/10

    N2 - Expression of the whey acidic protein (WAP) gene is tightly regulated in a tissue and developmental stage specific manner, in that the WAP gene is exclusively expressed in the mammary gland during pregnancy and lactation. Using both deletion and competition analyses, evidence is provided for the existence of a negative regulatory element (NRE) in the WAP promoter located between -413 and -93 with respect to the WAP transcriptional initiation site. This NRE dramatically decreases transcription from linked heterologous promoter-reporter gene constructs. The activity of NRE requires WAP promoter sequences that are 230 bp apart since subfragments of the NRE fail to inhibit transcription of adjoining reporter genes. Nuclear extracts from different cell types, in which the WAP gene is not active, contain a protein or complex that specifically interacts with the entire NRE but not with subfragments of it. The contact points between this protein (NRE binding factor [NBF]) and the NRE element have been partially determined. Mutation of the implicated nucleotides severely reduces the ability of NBF to bind, and such mutated promoter fragments fail to alleviate transcriptional repression in competition experiments. This suggests that NBF binding to the NRE is at least in part responsible for the negative regulation of the WAP promoter. Since NBF is not detectable in the lactating mammary gland, where the WAP gene is expressed, we speculate that it may be a determinant of the expression spectrum of the WAP gene. (C) 1994 Wiley-Liss, Inc.

    AB - Expression of the whey acidic protein (WAP) gene is tightly regulated in a tissue and developmental stage specific manner, in that the WAP gene is exclusively expressed in the mammary gland during pregnancy and lactation. Using both deletion and competition analyses, evidence is provided for the existence of a negative regulatory element (NRE) in the WAP promoter located between -413 and -93 with respect to the WAP transcriptional initiation site. This NRE dramatically decreases transcription from linked heterologous promoter-reporter gene constructs. The activity of NRE requires WAP promoter sequences that are 230 bp apart since subfragments of the NRE fail to inhibit transcription of adjoining reporter genes. Nuclear extracts from different cell types, in which the WAP gene is not active, contain a protein or complex that specifically interacts with the entire NRE but not with subfragments of it. The contact points between this protein (NRE binding factor [NBF]) and the NRE element have been partially determined. Mutation of the implicated nucleotides severely reduces the ability of NBF to bind, and such mutated promoter fragments fail to alleviate transcriptional repression in competition experiments. This suggests that NBF binding to the NRE is at least in part responsible for the negative regulation of the WAP promoter. Since NBF is not detectable in the lactating mammary gland, where the WAP gene is expressed, we speculate that it may be a determinant of the expression spectrum of the WAP gene. (C) 1994 Wiley-Liss, Inc.

    KW - negative regulatory element

    KW - heterologous promoter

    KW - DNA binding factor

    KW - transcriptional repression

    KW - milk protein expression control

    KW - cell-specific enhancer

    KW - high-level expression

    KW - long terminal repeat

    KW - tumor virus

    KW - transgenic mice

    KW - gene-expression

    KW - nuclear factor

    KW - hormonal-regulation

    KW - upstream sequences

    M3 - Article

    VL - 56

    SP - 245

    EP - 261

    JO - Journal of Cellular Biochemistry

    JF - Journal of Cellular Biochemistry

    SN - 0730-2312

    IS - 2

    ER -