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Related Experiment Videos

Multiple protein-DNA interactions within the human interferon-beta regulatory element.

S Xanthoudakis1, L Cohen, J Hiscott

  • 1Lady Davis Institute for Medical Research, Sir Mortimer B. Davis-Jewish General Hospital, Montreal, Quebec, Canada.

The Journal of Biological Chemistry
|January 15, 1989
PubMed
Summary

Researchers identified specific DNA-binding proteins interacting with the human interferon-beta regulatory element (IRE) in HeLa cells. These proteins are crucial for regulating interferon-beta gene expression, even in uninduced cells.

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Area of Science:

  • Molecular Biology
  • Virology
  • Gene Regulation

Background:

  • The human interferon-beta regulatory element (IRE) is critical for inducible gene expression.
  • Understanding the transcription factors that bind to the IRE is essential for deciphering interferon gene regulation.
  • HeLa S3 cells provide a model system for studying nuclear protein-DNA interactions.

Purpose of the Study:

  • To identify and characterize DNA-binding proteins specific for the human interferon-beta regulatory element (IRE).
  • To determine the precise DNA sequences within the IRE that interact with these proteins.
  • To investigate the role of these protein-DNA interactions in interferon-beta gene expression.

Main Methods:

  • Preparation of nuclear protein extracts from uninduced and induced HeLa S3 cells.

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  • Gel electrophoresis DNA binding assays to analyze protein-DNA interactions with an IRE probe.
  • Competition studies using various unlabeled DNA fragments to map binding sites and identify specific motifs.

    • Main Results:

    • Three specific DNA-protein complexes (A, B, and C) were identified for the IRE (-79 to -36).
    • The primary DNA binding domain was localized to the -79 to -64 region (P1 probe), essential for virus-inducible expression.
    • Adjacent upstream sequences (-94 to -78) also contained a motif binding the P1 protein, and IRE fragments inhibited in vitro transcription.

    Conclusions:

    • Multiple DNA sequence motifs within the interferon-beta regulatory element bind transcription regulatory proteins.
    • These regulatory proteins are present in both normal and virus-infected HeLa cells.
    • The findings elucidate key molecular mechanisms governing interferon-beta gene activation.