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Related Experiment Videos

Common double- and single-stranded DNA binding factor for a sterol regulatory element.

H C Stark1, O Weinberger, J Weinberger

  • 1Department of Medicine, Columbia University, New York, NY 10032.

Proceedings of the National Academy of Sciences of the United States of America
|March 15, 1992
PubMed
Summary

A novel nuclear factor, sterol regulatory element-binding factor (SRE-BF), binds to the sterol regulatory element-1 (SRE-1) sequence. This binding is sequence-specific for both single- and double-stranded DNA, suggesting a key role in sterol-regulated gene transcription.

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Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biochemistry

Background:

  • Sterol regulation of gene expression is crucial for maintaining cellular cholesterol homeostasis.
  • The sterol regulatory element-1 (SRE-1) is a cis-acting DNA sequence identified in the promoters of key genes involved in cholesterol metabolism, such as the low-density lipoprotein receptor (LDLR) and HMG-CoA reductase.

Purpose of the Study:

  • To identify and characterize nuclear factors that bind to the SRE-1 sequence.
  • To elucidate the mechanism by which SRE-1 mediates sterol-regulated transcription.

Main Methods:

  • Isolation and purification of nuclear factors from Chinese hamster ovary (CHO) cell nuclear extracts.
  • Electrophoretic mobility shift assays (EMSAs) to assess DNA-protein binding.
  • Competition assays using double-stranded and single-stranded DNA oligonucleotides.

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  • Photochemical cross-linking experiments to determine the molecular mass of the binding factor.

    • Main Results:

    • A nuclear factor, designated SRE-BF (sterol regulatory element-binding factor), was identified that specifically binds to the SRE-1 octanucleotide sequence.
    • SRE-BF exhibits sequence-specific binding to both double-stranded and single-stranded SRE-1 DNA.
    • Photochemical cross-linking revealed a common protein factor of 45-49 kDa that recognizes both forms of SRE-1.
    • The binding specificity of SRE-BF to single-stranded SRE-1 correlates with its known in vivo function in sterol-responsive transcription.

    Conclusions:

    • SRE-BF is a novel transcription factor that binds to the SRE-1 element.
    • SRE-BF recognizes both single- and double-stranded SRE-1 DNA, suggesting a versatile role in gene regulation.
    • The molecular characteristics of SRE-BF support its function as a key mediator of sterol-regulated gene expression.