A DNA element that regulates expression of an endogenous retrovirus during F9 cell differentiation is E1A dependent

B. T. Lamb, K. Satyamoorthy, D. Solter, A. Basu, M. Q. Xu, R. Weinmann, C. C. Howe

    Research output: Contribution to journalArticle

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    Abstract

    The retinoic acid-induced differentiation of F9 cells into parietal endoderm-like cells activates transcription of the endogenous mouse retrovirus, the intracisternal A-particle (IAP). To investigate the elements that control IAP gene differentiation-specific expression, we used methylation interference, Southwestern (DNA-protein), and transient- transfection assays and identified the IAP-proximal enhancer (IPE) element that directs differentiation-specific expression. We find that the IPE is inactive in undifferentiated F9 cells and active in differentiated parietal endoderm-like PYS-2 cells. Three proteins of 40, 60, and 68 kDa bind to the sequence GAGTGAC located between nucleotides -53 and -47 within the IPE. The 40- and 68-kDa proteins from both the undifferentiated and differentiated cells exhibit similar DNA-binding activities. However, the 60-kDa protein from differentiated cells has greater binding activity than that from undifferentiated cells, suggesting a role for this protein in F9 differentiation-specific expression of the IAP gene. The IAP gene is negatively regulated by the adenovirus E1A proteins, and the E1A sequence responsible for repression is located at the N terminus, between amino acids 2 and 67. The DNA sequence that is the target of E1A repression also maps to the IPE element. Colocalization of the differentiation-specific and E1A- sensitive elements to the same protein-binding site within the IPE suggests that the E1A-like activity functions in F9 cells to repress IAP gene expression. Activation of the IAP gene may result when the E1A-like activity is lost or inactivated during F9 cell differentiation, followed by binding of the 60-kDa positive regulatory protein to the enhancer element.

    Original languageEnglish
    Pages (from-to)4824-4833
    Number of pages10
    JournalMolecular and Cellular Biology
    Volume12
    Issue number11
    Publication statusPublished - 1992

    Fingerprint

    Endogenous Retroviruses
    Intracisternal A-Particle Genes
    Cell Differentiation
    DNA
    Endoderm
    Proteins
    Adenovirus E1A Proteins
    Tretinoin
    Protein Binding
    Methylation
    Transfection
    Nucleotides
    Binding Sites
    Gene Expression
    Amino Acids

    All Science Journal Classification (ASJC) codes

    • Molecular Biology
    • Cell Biology

    Cite this

    Lamb, B. T. ; Satyamoorthy, K. ; Solter, D. ; Basu, A. ; Xu, M. Q. ; Weinmann, R. ; Howe, C. C. / A DNA element that regulates expression of an endogenous retrovirus during F9 cell differentiation is E1A dependent. In: Molecular and Cellular Biology. 1992 ; Vol. 12, No. 11. pp. 4824-4833.
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    abstract = "The retinoic acid-induced differentiation of F9 cells into parietal endoderm-like cells activates transcription of the endogenous mouse retrovirus, the intracisternal A-particle (IAP). To investigate the elements that control IAP gene differentiation-specific expression, we used methylation interference, Southwestern (DNA-protein), and transient- transfection assays and identified the IAP-proximal enhancer (IPE) element that directs differentiation-specific expression. We find that the IPE is inactive in undifferentiated F9 cells and active in differentiated parietal endoderm-like PYS-2 cells. Three proteins of 40, 60, and 68 kDa bind to the sequence GAGTGAC located between nucleotides -53 and -47 within the IPE. The 40- and 68-kDa proteins from both the undifferentiated and differentiated cells exhibit similar DNA-binding activities. However, the 60-kDa protein from differentiated cells has greater binding activity than that from undifferentiated cells, suggesting a role for this protein in F9 differentiation-specific expression of the IAP gene. The IAP gene is negatively regulated by the adenovirus E1A proteins, and the E1A sequence responsible for repression is located at the N terminus, between amino acids 2 and 67. The DNA sequence that is the target of E1A repression also maps to the IPE element. Colocalization of the differentiation-specific and E1A- sensitive elements to the same protein-binding site within the IPE suggests that the E1A-like activity functions in F9 cells to repress IAP gene expression. Activation of the IAP gene may result when the E1A-like activity is lost or inactivated during F9 cell differentiation, followed by binding of the 60-kDa positive regulatory protein to the enhancer element.",
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    A DNA element that regulates expression of an endogenous retrovirus during F9 cell differentiation is E1A dependent. / Lamb, B. T.; Satyamoorthy, K.; Solter, D.; Basu, A.; Xu, M. Q.; Weinmann, R.; Howe, C. C.

    In: Molecular and Cellular Biology, Vol. 12, No. 11, 1992, p. 4824-4833.

    Research output: Contribution to journalArticle

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    AU - Satyamoorthy, K.

    AU - Solter, D.

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    AU - Xu, M. Q.

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    AU - Howe, C. C.

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